JP4015227B2 - Chemical reaction processing apparatus and method using magnetic particles - Google Patents

Description

この数式には、どのように試薬を配列又は移動（転置移動を含む）して、どのような目的物質がどの位置にある容器に得られるかの情報が含まれている。
【００２１】
第二の発明は、第一の発明において、前記ピペット手段のノズルの配列、又は前記処理ステージの容器の配列は、相互に関連し、且つ、処理過程の内容に基づいて定められたものである。
【００２２】
ここで、ノズルの配列には、１列（行）状の場合のみならずマトリクス状に配列された場合も含む。「相互に関連する」例として、ノズルが各容器に接触せずに進入可能なように各径を定め、ノズル間の間隔と容器群間の間隔とが一致する場合やノズル間の間隔が容器群間の間隔が整数倍の場合や、ノズル群の配列の個数と容器群の行又は列の個数とを一致又はその整数倍又は整数分の一にさせたりすることを含む。また、「処理過程の内容」の例として、反応に必要な物質群の種類の個数分に等しい個数をもつ行数又は、別の物質群の種類の列数をもつマトリクス状に配列された容器群を用いる場合がある。
【００２３】
第三の発明は、第一の発明又は第二の発明において、前記ピペット手段は、液体吸引ラインの分注機のノズルに着脱自在に装着する分注チップを有するものである。
ここで、例えば、分注チップはプラスチックス、ステンレススチール又はテフロンによって形成される。
【００２４】
第四の発明は、第一の発明乃至第三の発明において、前記処理ステージの容器群の一部又は全体の各容器には、外部に磁力手段が設けられて各容器内面での磁性体粒子の吸着及び脱着が可能なものである。
これによって、前記制御手段は各容器内に収容された収容物のうちの移送対象として、結合体以外のもの又はその一部を移送するように制御可能である。
【００２５】
第五の発明は、第一の発明において、前記処理ステージに設けられた容器群の一部又は全体の各容器上が膜状又は板状の蓋部材によって覆われ、前記ピペット手段のノズルに装着された分注チップの先端によって貫通可能であるものである。
「貫通可能」であるためには、蓋部材に貫通用の孔が予め設けられている場合と、孔が予め設けられていない蓋部材を分注チップの先端で貫通する場合がある。このような蓋部材としては、ラミネートやゴム板等がある。
【００２６】
第六の発明は、第一の発明乃至第五の発明において、前記処理ステージには分注用の容器、洗浄用の容器、反応用の容器、恒温装置又は／及び測定用装置等の処理過程に必要な器具が設けられているものである。
【００２７】
第七の発明は、第一の発明乃至第六の発明のいずれかにおいて、前記処理ステージ及び前記ピペット手段は密閉容器内に設けられたものである。
第八の発明は、第一の発明乃至第七の発明において、前記制御手段は、前記ピペット手段、前記処理ステージ又は前記移動手段に対して、処理過程中の処理時間、若しくは、前記移動手段による移動量、移動時間若しくは消費エネルギーを小さくし、又は動作を単純化することによって効率良く制御するものである。
【００２８】
第九の発明は、第一の発明乃至第八の発明に係るピペット手段、処理ステージ、指定手段、移動手段及び制御手段を用いることによって、指定された処理過程の内容に基づいて、配列された前記処理ステージの容器群の一部又は全体に複数種類の試薬等の物質群をマトリクス状に配置する過程（Ｓ１）と、指定された処理過程の内容に基づいて、物質群が配置された前記容器群の一部又は全体に磁性体粒子を含む液を分注して、各物質中の目的物質と磁性体粒子とを反応等によって結合させて結合体を生成する過程（Ｓ２）と、指定された処理過程の内容に基づいて、ピペット手段のノズル群又は処理ステージの容器群の一部又は全体について行と列を入れ換えて移動する転置移動又は行と列とを入れ換えずに移動する並進移動によって、移送元の容器又は容器群に収容された収容物のうちの一部又は全体を、移送先の容器又は容器群に移送する過程（Ｓ３）とを含むものである。
【００２９】
ここで、収容物のうち結合体のみを移送するには、前記ピペット手段に設けた磁力手段がピペット手段の液通路に磁場を及ぼすことによって磁性体粒子を分離して、内壁に保持することによって行う。
結合体以外のものを移送する場合には、例えば、容器に磁性体を設けることによって結合体を容器の内壁に吸着保持させることによって行うことができる。
【００３０】
第十の発明は、第九の発明において、前記過程（Ｓ１）は、マトリクス状に配列された容器群のうちａ₁ 〜ａ_n の試薬を等量ずつｎ列×１行の容器群に分注及び配置し、ｂ₁ 〜ｂ_m の試薬を等量ずつｍ列×１行の容器群に分注及び配置する過程であり、前記過程（Ｓ２）は試薬が配置された前記容器群に磁性体粒子を分注して、ａ₁ 〜ａ_n の試薬中の目的物質と磁性体粒子とを反応等によって結合させて結合体を生成する過程であり、前記過程（Ｓ３）は、前記結合体が含まれた各容器の収容物全体をピペット手段によって共通の混合容器内に移送して混合し、該混合液をｍ等分し、ｎ列×ｍ行のマトリクス状に配列された容器の各列ｍ行の容器群に等量ずつピペット手段によって移送するとともに、該移送の際に並進移動又は転置移動を選択して、ｂ₁ 〜ｂ_m の試薬をｎ等分し、前記ｎ列×ｍ行のマトリクス状に配列された容器の各列ｎ行の容器群に等量ずつピペット手段によって移送する過程であるものである。
ここで、ｎ＝ｍの場合には、ｍ種の試薬を最小回数で漏れなく分注することができるので、特に転置移動の必要はない。転置移動にするか並進移動にするかは、その他ピペット手段のノズル群の配列にも依存する。
本発明はいわゆるスプリット・プール法により化学反応処理を行うものである。
【００３１】
第十一の発明は、第九の発明において、前記過程（Ｓ１）は、マトリクス状に配列された容器群のうちａ₁ 〜ａ_n の試薬を等量ずつ、ｎ列×ｍ行の容器群に等量ずつ分注及び配置し、ｂ₁ 〜ｂ_m の試薬を等量ずつｍ列×１行の容器群に分注及び配置する過程であり、前記過程（Ｓ２）は、ｎ列×ｍ行のマトリクス状に配列された全容器に各々等量の磁性体粒子を分注して前記試薬中の目的物質と磁性体粒子とを結合させる過程であり、前記過程（Ｓ３）は、前記ｎ列×ｍ行のマトリクス状に配列された全容器に、該容器の外部に設けられた磁力手段によって磁場を及ぼし、磁性体粒子との結合体を各容器の内面に吸着保持させ、前記ピペット手段によって、容器内面に吸着保持されている結合体以外の収容物を吸引して取り除き、前記ｎ列×ｍ行の容器群又はピペット手段のノズル群を転置移動することによって、前記結合体が収容された前記ｎ列×ｍ行のマトリクス状に配列された全容器に、前記ｂ₁ 〜ｂ_m の各容器に収容されている試薬をｎ等分して分割して移送する過程である。
本発明は、磁力手段を処理ステージの容器の容器側に配設した場合に行うパラレル合成法を示したものである。
【００３２】
第十二の発明は、第九の発明において、前記過程（Ｓ１）は、ｎ列×ｍ行のマトリクス状に配列されたｍ／ｎ群個の各ｎ列×ｍ行容器群に、a₁〜 a_n のｎ個の各種類の試薬を、各列のｍ個の容器群に等量ずつ分注して配置する過程であり、前記過程（Ｓ２）は、ｍ／ｎ群個の各ｎ列×ｍ行容器群のうち、第１群のｎ列×ｍ行容器群の全容器に各々等量の磁性体粒子を分注して前記各試薬に含まれる目的物質と磁性体粒子とを結合させて結合体を生成する過程であり、前記過程（Ｓ３）は、各ｎ列×ｍ行容器群の各容器の収容物のうち前記結合体のみを磁力手段を配設したピペット手段を用いて、第１群のｎ列×ｍ行容器群から第ｍ／ｎ群のｎ列×ｍ行容器群に達するまで順次、転置移動若しくは並進移動によって、次群のｎ列×ｍ行容器群に移送する過程である。
【００３３】
本発明は、ピペット手段に磁力手段を配設した場合のパラレル合成法に対応するものである。
「ｍ／ｎ（＝ｋ）」は、最終的な生成物に含まれる成分ａ_i （ここでは、各試薬に含まれる目的物質を表すとすると）の結合の数であり、最終的な生成物がａ_i1−ａ_i2−…−ａ_ik（i1,i2,…,ik=1,2, …n)のような構成になることを意味している。
例えば、最終目的生成物がａ_i −ａ_j −ａ_k のような３成分系の場合には、３個のｎ列×ｍ行容器群を予め処理ステージに用意しておくことになる。
【００３４】
第十三の発明は、第九の発明乃至第十二の発明のいずれかにおいて、物質群の各物質とともに、該各物質を識別するための識別子を添加して、該識別子を磁性体粒子に結合又は磁性体粒子の周辺に位置させることによって、磁性体粒子に結合している物質を識別するものである。
ここで、「識別子」には、タグのみならず、情報記憶用のチップも含む。
「周辺に位置する」には、浸透性のあるカプセル内に閉じ込められる場合も含む。
【００３５】
【発明の実施の形態】
図１〜図６に基づいて、本発明の第一の実施の形態について説明する。
図１に、本実施の形態に係る磁性体粒子を用いた化学処理装置をコンビナトリアル合成処理に適用した場合を示す。同図に示すように、本実施の形態に係る装置は、液体の吸引及び吐出等を行うマルチノズルを有するピペット手段１と、液体を収容する容器群がｘｙ軸に沿ってマトリクス状に配列されたプレートを少なくとも有する処理ステージ２と、前記ピペット手段１の各ノズルに連通する液通路又は各処理ステージ２の各容器内面での磁性体粒子の吸着及び脱着可能な磁力手段（図示せず）と、前記ピペット手段１について前記処理ステージ２の盤面に沿ったｘｙ方向及びｚ方向に沿ってノズル群と容器群の行と列を入れ換えずに移動する並進移動及び行と列を入れ換えて移動する転置移動をさせ、並びに前記処理ステージ２についてその一部容器群をその面内において行と列を入れ換える転置をさせる回動可能なモータ等からなる移動手段３とを有する。
【００３６】
また、本化学処理装置は、コンビナトリアル合成等の化学反応処理の内容を指定する指定手段に相当する入力手段６と、前記ピペット手段１の吸引及び吐出の動作、前記磁力手段を用いた磁性体粒子の吸着及び離脱等の動作、前記処理ステージの容器群に対する関与物質群の配置、各容器内に収容された収容物のうちの移送対象、並びに、移送元及び移送先の容器若しくは容器群、移送の際の行と列とを入れ換えて移動する転置移動若しくは行と列とを入れ換えずに移動する並進移動の別を、前記入力手段６等で指定された処理過程の内容に基づいて、特定して各手段に指示するＣＰＵ等からなる制御手段４とを有する。
【００３７】
さらに、本装置は、処理過程である反応過程の設定や、反応過程の経過等の種々の表示を画面に行う表示手段５と、キーボード、マウス若しくはスイッチの操作又はフロッピー若しくはＣＤの装着等によって種々のコンビナトリアル合成等の指定を行う入力手段６と、各種試薬の分注量、分注位置、分注のタイミング等の種々のデータを記憶する記憶手段７と、データの伝送を行う通信手段８とを有する。これらの各装置及び手段はバス等を介して相互に接続されている。
尚、前記入力手段６及び記憶手段７は前記指定手段に相当するものである。
【００３８】
図２には、前記処理ステージ２の上面図である。図中、符号１０は、前記ピペット手段１に設けられた６連のノズル群であり着脱自在の分注チップが装着されている。該ノズルには、液面の検知を行うために光を照射して、その反射光を受光する液面センサ（図示せず）や、ノズル内の圧力を検知する圧力センサが設けられている。
尚、用いられる化学反応が環境に悪影響を生じさせるような場合や、空気以外の気体や真空等の種々の雰囲気下で処理を行う場合には、前記ピペット手段及び処理ステージを、密閉容器９に収納して密閉する。
【００３９】
本処理ステージ２は、ノズル群１０が同一の試薬を分注する場合に用いるノズル群１０に装着された分注チップが進入する部分が相互に仕切られていない同一分注試薬用容器１１、１３が設けられている。符号１３の同一分注試薬用容器の場合は、符号１１の場合よりも、大きな容量の試薬を収容することができる。どちらを用いるかは目的化合物群の量等の処理過程の内容に基づいて選択される。
【００４０】
処理ステージ２は前記６連のノズル群１０が各々異なる試薬を分注する場合に用いる、ノズル群１０の各ノズルに装着された分注チップが進入する部分が相互に仕切られている異分注試薬用容器１２，１４が設けられている。符号１４の異分注試薬用容器の場合は、符号１２の容器よりも容量が大きい場合に選択される。
【００４１】
該処理ステージ２は、前記ノズル群１０に装着され且つ取り外された分注チップが保持されているチップホルダ１５と、試薬分注を行った分注ノズルを洗浄するための洗浄液が収容されている試薬分注ノズル洗浄槽１６と、反応した化合物のノズルを洗浄するための洗浄液が収容されている反応ノズル洗浄槽１７と、６行×６列のマトリクス状に容器群が配列され、反応が行われる反応用プレート１８と、試薬及び磁性体粒子を混合するための混合用プレート１９とを有する。
【００４２】
前記反応用プレート１８、混合用プレート１９は各々独立して９０°の角度の正逆両方向への回転、即ち、行と列を入れ換える転置が可能な回転ステージ２０上に設けられている。該回転ステージ２０は前記移動手段３の１つに相当する。
【００４３】
前記同一分注試薬用容器１１、１３及び前記異分注試薬用容器１２，１４は前記分注チップが進入する部分を除いて容器の開口部が蓋体で覆われている。これによって、試薬の蒸発や汚染を防止している。一方、試薬分注ノズル洗浄槽１６、反応ノズル洗浄槽１７、反応用プレート１８、混合用プレート１９には蓋体が設けられていない。
【００４４】
好ましくは、処理ステージ２のうち容器群の部分をラミネートシーリング膜で覆うことによって、容器群を密閉し、例えば窒素等を充満させた雰囲気を保ち、試薬の蒸発や汚染を確実に防止することができる。ラミネートシーリング膜を用いた場合には、該膜に穴を開けるピアシング方式を採用する。前記分注チップは、例えば、ポリプロピレン（ＰＰ）、ステンレススチール（ＳＵＳ）やテフロンによって形成し、ピアッシング方式を採用する場合には針状に形成するのが適当である。
【００４５】
図３は、図２に示した処理ステージ２に着脱可能に設けられた同一分注試薬用容器１１を拡大して示した分解透視斜視図である。該同一分注試薬用容器１１は、ＰＰで形成された箱状の容器部１１ａと、該容器部１１ａの開口部１１ｂに着脱自在に嵌合して覆うＰＰで形成された略長方形状の蓋体１１ｃを有する。
【００４６】
該容器部１１ａは側面部１１ｄと、内底部１１ｇとから形成される。ここで、内底部１１ｇは、該内底部１１ｇの中央に向かって落ち込む傾斜面１１ｅと、前記分注チップ（図示せず）の先端部の口径よりも幅狭で該内底部１１ｇの中央に沿って設けられた溝部１１ｆとを有する。
【００４７】
これによって、該容器１１の内底面に分注チップの先端部が当接しても該容器内の試薬をほぼ全量吸引することができるとともに、容器内に吸引した試薬を吐出するときの試薬の拡散を平均化して攪拌効率を大幅に向上することができる。長手方向にある前記側面部１１ｄの側面１１ｅは該容器１１が倒れないように安定して支持するために、その下端が水平となるように長方形状に形成され、上端には、容器を把持するたの把手１１ｈが設けられている。
【００４８】
蓋体１１ｃの中央には略長方形状の窪みが設けられ、該窪みには十字状や細孔状の切込み１１ｉが６個、所定の間隔で設けられている。これは６連、３連、２連又は単独の分注チップが設けられたピペット手段１に対応し、該分注チップが該切込み１１ｉを貫通して容器内部に進入することができる。この切込み１１ｉを設けることによって、分注チップの液を拭い、容器内の液体の蒸発を防止し、且つ汚染防止に役立つ。試薬を保存する場合には、完全に液体の蒸発を防止するためゴム板１１ｊが前記蓋体１１ｃの中央の前記窪みに着脱自在に嵌め込まれる。
尚、前記切込みや細孔をゴム板１１ｊに設けるようにしても、設けずに分注チップによって貫通可能とするようにしても良い。
【００４９】
図４は、図３に示した同一分注試薬用容器１１よりも容量の小さい同一分注試薬用容器２１を示す。該容器２１は、ＰＰで形成された細長い箱状の容器部２１ａと、該容器部２１ａの開口部２１ｂに着脱自在に嵌合して覆うＰＰで形成された細長い長方形状の蓋体２１ｃとを有する。該容器部２１ａは側面部２１ｄと、内底部２１ｇとから形成される。ここで、該内底部２１ｇは、くさび底であり、該内底部２１ｇの中央に向かって落ち込むように傾斜する傾斜面２１ｅと、前記分注チップ（図示せず）の先端部の口径よりも幅狭で該内底部２１ｇの中央に沿って設けられた溝部２１ｆとを有する。
【００５０】
溝部２１ｆの底面は、複数個（この例では１２個）の円弧状の凹部がつながった波形の曲面が形成されている。これによって、該容器２１の内底面に分注チップの先端部が当接しても該容器内の試薬をほぼ全量吸引することができるとともに、容器内に吸引した試薬を吐出するときの試薬の拡散を平均化して攪拌効率を大幅に向上することができる。長手方向にある前記側面部２１ｄの側面は該容器２１が倒れないように安定して支持するために、その下端が水平となるように長方形状に形成され、上端には、容器を把持するための把手２１ｈが設けられている。
【００５１】
前記蓋体２１ｃの中央には略長方形状の窪みが設けられ、該窪みには十字状や細孔状の切込み２１ｉが１２個、前記溝部２１ｆの各円弧状の凹部に対応する位置に設けられている。これは１２連、６連、４連、３連、２連又は単独の分注チップが設けられたピペット手段１に対応し、該分注チップが該切込み２１ｉを貫通して容器内部に進入することができる。
【００５２】
この切込み２１ｉを設けることによって、分注チップの挿入の際に外部に付着されている液を拭いクロスコンタミネーションを防止するとともに、容器内の液体の蒸発を防止し、ゴミ等の侵入等による汚染防止に役立つ。
試薬を保存する場合には、完全に液体の蒸発を防止するためゴム板２１ｊが前記蓋体２１ｃの中央の前記窪みに着脱自在に嵌め込まれる。
尚、該ゴム板２１ｊにも切込みや細孔を設けるようにしても、また設けずに分注チップにより貫通可能とするようにしても良い。
【００５３】
図５は、図４に示した同一分注試薬用容器２１の変形例を示すものである。
該同一分注試薬用容器３１は、ＰＰで形成された細長い箱状の容器部３１ａと、該容器部３１ａの開口部３１ｂに着脱自在に嵌合して覆うＰＰで形成された細長い長方形状の蓋体３１ｃとを有する。該容器部３１ａと蓋体３１ｃとは薄肉に形成されたヒンジ３１ｄによって屈曲自在に結合され、ヒンジ３１ｄが折り曲げらると、蓋体３１ｃが前記開口部３１ｂに嵌合するように着脱自在に形成されている。
【００５４】
該容器部３１ａは側面部３１ｅと、内底部３１ｈとから形成されている。ここで、該内底部３１ｈは、くさび底をしており該内底部３１ｈの中央に沿って、下方に向かって狭まるように傾斜する傾斜面３１ｆと、該内底部３１ｈの中央に沿って前記分注チップ（図示せず）の先端部の口径よりも幅狭で設けられた溝部３１ｇとを有する。
【００５５】
溝部３１ｇの底面は、複数個（この例では１２個）の凹部をもつジグザグの波形の面が形成されている。これによって、該容器３１の内底面に分注チップの先端部が当接しても該容器内の試薬をほぼ全量吸引することができるとともに、容器内に吸引した試薬を吐出するときの試薬の拡散を平均化して攪拌効率を大幅に向上することができる。
【００５６】
前記容器部３１ａの上端の前記開口部３１ｂの周縁には、前記容器部３１ａの側面部３１ｅよりも大きめで特に長手方向の両端で把持可能な大きさの板が突出するように形成された上板３１ｉが設けられている。該上板３１ｉの長手方向に突出した部分の下面で側面部３１ｅの側面から突出した支持部３１ｊにより支えられている。
【００５７】
前記蓋体３１ｃの中央に細長い長方形状の孔を穿設し、該孔にはゴム板３１ｋが嵌められている。前記蓋体３１ｃに嵌められたゴム板３１ｊを貫通して分注チップが挿入されるように十字の切込み３１ｌが１２個、前記溝部３１ｇの底面の１２個の波形の凹部に対応する位置に設けられている。これは１２連、６連、４連、３連、２連、又は単独の分注チップが設けられたピペット手段１に対応するものである。
尚、以上説明した容器の蓋に設けたゴム板に切込みを設けずに、ピペット手段の先端によって、孔を開けて貫通するようにしても、貫通不可能とするようにしても良い。この場合には切込みがないので容器内部は完全に密閉されることになる。
【００５８】
本実施の形態によれば、蓋体が設けられているので、蓋体と容器とが別々になる事態を防止し、したがって、クロスコンタミネーションを防止することになる。
【００５９】
図６（ａ）は前記処理ステージ２に設けられた分注チップ保持部１５の断面を示すものであり、同図（ｂ）は、前記洗浄槽１６，１７の断面図を示す。前記洗浄槽１６，１７は各々分注ピペットの吸引吐出が行われる洗浄皿１６ａと、該洗浄皿１６ａから溢れた洗浄液を回収するための受け皿１６ｂとを有する。
【００６０】
さらに、同図に示すように、該洗浄皿１６ａに洗浄液を供給するための供給路１６ｃ及びポンプ１６ｄと、洗浄液を貯溜する貯溜槽１６ｅと、前記受け皿１６ｂに設けられた洗浄液を廃棄する廃棄路１６ｆとが設けられている。
前記処理ステージ２には、その他、試薬を保存し、又は反応を促進するための恒温装置や、液面を検知するための液面センサを設けるようにしても良い。
【００６１】
続いて、図７乃至図１１に基づいて、第一の実施の形態に係る化学反応処理装置を用いて、コンビナトリアル合成及び構造解析を行う種々の場合について、以下に説明する。
最初に、第二の実施の形態として、図７及び図８に基づいて、スプリット合成法(split synthesis) により、３種類のアミノ酸（Ａ，Ｂ，Ｃ）を使用して、トリペプチドを最終生成化合物として合成する例について説明する。この場合には、前記磁力手段が、少なくとも、ピペット手段１の各ノズルと連通する液通路の外部に配設されているとする。
【００６２】
図７に示すように、ステップＳ１で、前記入力手段６から、コンビナトリアル合成の処理過程の内容を特定するため、処理過程情報を入力する。処理過程を特定すると、スプリット合成を行う点、試薬が収容された容器の位置、磁性体粒子が収容された容器の位置、反応が行われる容器の位置、ピペット手段又は処理ステージの移動の制御、反応時間、分注の順序等についても、予め前記記憶手段７に格納された情報に基づいて自動的に特定又は設定される。これによって、必要な試薬の配列等が前記制御手段４によって決定されることになる。
【００６３】
ステップＳ２で、処理過程がスプリット合成なので、自動的にステップＳ３に進む。ステップＳ３で、前記ピペット手段１のノズル群１０が、前記処理ステージ２のチップホルダ１５に移動して６連の分注チップのうち３連分を自動的に装着し、処理ステージ２の異分注試薬用容器１２にノズル群１０を移動させて、該容器１２に収容されている目的物質であるアミノ酸Ａ，Ｂ，Ｃを含む試薬を各々吸引して、図８に示すように、反応用プレート１８₁ の６行×６列のマトリクス状に配列された容器群のうち３行×１列分の容器群に前記アミノ酸を吐出して配置する。
【００６４】
ステップＳ４で、分注チップが装着された前記ノズル群１０は、同一分注試薬用容器１１に移動し、磁性体粒子が貯溜されている該容器１１から磁性体粒子の懸濁液を吸引する。ここで、磁性体粒子の表面には、前記アミノ酸Ａ，Ｂ，Ｃと特異的に反応する物質がコーティング又は保持されている。その際、アミノ酸Ａ，Ｂ，Ｃに応じて、コーティングされている物質の異なる複数種類の磁性体粒子を用いても良い。
【００６５】
次に、前記アミノ酸Ａ，Ｂ，Ｃが各々収容されているプレートの３行×１列のウェルに移動し、磁性体粒子をアミノ酸Ａ，Ｂ，Ｃを含む試薬が収容されている各容器に分注して混合させて、各アミノ酸Ａ，Ｂ，Ｃを磁性体粒子と反応させて結合させる。この段階で、前記３行×１列の各容器には、磁性体粒子−Ａ、磁性体粒子−Ｂ、磁性体粒子−Ｃの各結合体が収容されることになる。
【００６６】
ステップＳ５で、図８の反応用プレート１８₁ の３個の容器に収容された各アミノ酸Ａ，Ｂ，Ｃと磁性体粒子との結合体を含む混合液は、分注チップが装着された前記ノズル群１０によって吸引され、移動手段３によって前記処理ステージ２の混合用プレート１９に移動して、各混合液を一斉に吐出して、磁性体粒子と各アミノ酸Ａ，Ｂ，Ｃとの結合体との懸濁液を混合してランダム化し、前記結合体が等量ずつ一様に混ざった混合液を生成する。
【００６７】
ステップＳ６で、該混合用プレート１９で混合されたアミノ酸Ａ，Ｂ，Ｃと磁性体粒子との結合体が一様に懸濁した該混合液を、再び３連の前記ノズル群１０によって等量ずつ吸引し、吸引した各混合液を移動手段３によって、図８に示すように、前記反応用プレート１８₁ の６列×６行のマトリクスの部分マトリクスである３列×３行の容器群９個に並進移動しつつ、各ノズル毎に吸引した混合液の１／３（最初の各アミノ酸Ａ，Ｂ，Ｃの量を１とした場合に）ずつを、計９個の容器群に、等分に分配する。その際、行と列との数が３で同じなので該プレートは９０°回転して転置する必要はない。
【００６８】
ステップＳ７で、前記ノズル群１０は、再び、前記異分注試薬用容器１２に各々収容されている前記アミノ酸Ａ，Ｂ，Ｃが別々に含まれている各試薬各々１ずつ吸引して、３行×３列の前記マトリクス行列の各要素に１／３ずつ、各ノズルは１列×３行の容器群に並進移動することによって列毎に異なるアミノ酸を等量ずつ分割して分注する。
【００６９】
したがって、図８の３行×３列のマトリクスの各容器には各々アミノ酸Ａ，Ｂ，Ｃを含む試薬と磁性体粒子とアミノ酸Ａ，Ｂ、Ｃとの結合体の混合液とが混合されることになる。
【００７０】
ステップＳ８で、アミノ酸同士の反応及びインキュベーションによって、２種のアミノ酸が結合した結合体が生成される。
したがって、第１列の容器群では、磁性体粒子−Ａ−Ａ、磁性体粒子−Ｂ−Ａ、磁性体粒子−Ｃ−Ａ、第２列の容器群では、磁性体粒子−Ａ−Ｂ、磁性体粒子−Ｂ−Ｂ、磁性体粒子−Ｃ−Ｂ、第３列の容器群では、磁性体粒子−Ａ−Ｃ、磁性体粒子−Ｂ−Ｃ、磁性体粒子−Ｃ−Ｃという各結合体からなる化合物群が得られることになる。
【００７１】
ステップＳ９で、前記ノズル群は３列×３行のマトリクス状の容器群から、各行毎に順次、各収容物を全量吸引して、前記混合用プレート１９の１つの容器に吐出し、前記化合物群の一様な混合液を生成する。該混合液には前記各結合体が均一に混合する。
【００７２】
ステップＳ１０で、前記ノズル群１０は、該混合用プレート１９から、等量ずつ９回に分けて混合液を吸引し、図８の反応用プレート１８₁ ，１８₂ のうち各々３列×６行及び３列×３行のマトリクスの各容器群に、前記ノズル群１０を並行移動させることによって、各行毎にアミノ酸Ａ，Ｂ，Ｃの結合体を含む混合液の全量の１／２７ずつ、即ち２／９ずつ等量分割して分注する。
【００７３】
ステップＳ１１で、前記ノズル群１０は、前記異分注試薬用容器１２から、さらに、各々１ずつアミノ酸Ａ，Ｂ，Ｃを各ノズル毎に吸引し、該ノズル群１０を各行毎に並進移動しつつ、１／９ずつ等量に、分割分注して、各ウェルに収容されていた混合液と混合する。この場合も、回転ステージ２０₁ ，２０₂ を９０°回転して転置させる必要はない。
【００７４】
ステップＳ１２で、アミノ酸同士の反応及びインキュベーションによって、３種のアミノ酸が結合した結合体が生成される。これによって、３列×６行の各ウェルには、各列毎に、次のような結合体が含まれることになる。第１列については、磁性体粒子−Ａ−Ａ−Ａ、磁性体粒子−Ｂ−Ａ−Ａ、磁性体粒子−Ｃ−Ａ−Ａ、磁性体粒子−Ａ−Ｂ−Ａ、磁性体粒子−Ｂ−Ｂ−Ａ、磁性体粒子−Ｃ−Ｂ−Ａ、磁性体粒子−Ａ−Ｃ−Ａ、磁性体粒子−Ｂ−Ｃ−Ａ、磁性体粒子−Ｃ−Ｃ−Ａである。
【００７５】
第２列については、磁性体粒子−Ａ−Ａ−Ｂ、磁性体粒子−Ｂ−Ａ−Ｂ、磁性体粒子−Ｃ−Ａ−Ｂ、磁性体粒子−Ａ−Ｂ−Ｂ、磁性体粒子−Ｂ−Ｂ−Ｂ、磁性体粒子−Ｃ−Ｂ−Ｂ、磁性体粒子−Ａ−Ｃ−Ｂ、磁性体粒子−Ｂ−Ｃ−Ｂ、磁性体粒子−Ｃ−Ｃ−Ｂである。
【００７６】
第３列については、磁性体粒子−Ａ−Ａ−Ｃ、磁性体粒子−Ｂ−Ａ−Ｃ、磁性体粒子−Ｃ−Ａ−Ｃ、磁性体粒子−Ａ−Ｂ−Ｃ、磁性体粒子−Ｂ−Ｂ−Ｃ、磁性体粒子−Ｃ−Ｂ−Ｃ、磁性体粒子−Ａ−Ｃ−Ｃ、磁性体粒子−Ｂ−Ｃ−Ｃ、磁性体粒子−Ｃ−Ｃ−Ｃである。この段階で、全２７種類のトリペプチドが磁性体粒子と結合した結合体が得られることになる。
【００７７】
ステップＳ１３で、該３列×６行、及び３列×３行のマトリクス状に配列された容器群であるウェル群に解離剤を分注して、各々前記結合体を磁性体粒子から解離させ、磁性体粒子のみを分離することによって、各トリペプチドが得られることになる。
【００７８】
続いて、第三の実施の形態として、図７及び図９に基づいて、Ａ_i −Ｂ_j −Ｃ_k という最終生成化合物を、少なくとも、磁力手段（図示せず）が処理ステージ２の各容器（反応用）の外部に各々配設され、各容器内面で磁性体粒子の吸着及び脱着が可能な装置を用いた場合にパラレル合成法を適用する場合を説明する。
【００７９】
図７のステップＳ１で、操作者は、図１の前記入力手段６から、コンビナトリアル合成の処理過程を特定するために処理過程情報を入力する。処理過程を特定すると、パラレル合成を行う点、その試薬の収納容器位置、磁性体粒子の収納容器位置、反応が行われる容器位置、ピペット手段又は処理ステージの移動の制御、反応時間、分注の順序等についても、予め前記記憶手段７に格納された情報に基づいて自動的に特定又は設定される。
【００８０】
ステップＳ２で、処理過程が、パラレル合成法なので、自動的にステップＳ２１に進む。
ステップＳ２１で、前記ピペット手段１のノズル群１０が、前記処理ステージ２のチップホルダ１５に移動して６連の分注チップのうち３連分を自動的に装着し、前記処理ステージ２の異分注試薬用容器１２にノズル群１０を移動させて、該容器１２に収容されている目的物質Ａ₁ ，Ａ₂ ，Ａ₃ が別々に含まれている各試薬をノズル群１０の各ノズルが独立に吸引して、図９に示すように、反応用プレート１８₁ の６行×６列のマトリクス状に配置された容器群のうち３行×１列分の容器群に前記試薬を吐出する。
【００８１】
ステップＳ２２で、分注チップが装着された前記ノズル群１０は、同一分注試薬用容器１１に移動し、磁性体粒子が貯溜されている該容器１１から磁性体粒子を含む懸濁液を吸引する。ここで、磁性体粒子の表面には、前記目的物質Ａ₁ ，Ａ₂ ，Ａ₃ と特異的に反応する物質がコーティングされている。その際、目的物質Ａ₁ ，Ａ₂ ，Ａ₃ に応じて、コーティングされている物質の異なる複数種類の磁性体粒子を用いても良い。
【００８２】
次に、前記目的物質Ａ₁ ，Ａ₂ ，Ａ₃ が各々収容されているプレートの３行×１列のウェルに移動し、磁性体粒子を該目的物質Ａ₁ ，Ａ₂ ，Ａ₃ が別々に含まれている各試薬が収容されている各容器に分注して、各目的物質Ａ₁ ，Ａ₂ ，Ａ₃ を磁性体粒子と結合させて磁性体粒子と結合させる。この段階で、該３行×１列の容器群には、順次、磁性体粒子−Ａ₁ 、磁性体粒子−Ａ₂ 、磁性体粒子−Ａ₃ の各結合体を含む液が収容されていることになる。
【００８３】
ステップＳ２３で、前記ノズル群１０は、磁性体粒子と結合した結合体を含む各試薬を吸引して、当初の量の１／３ずつ、並進移動しながら、等量に吐出することによって、反応用プレート１８₁ の６行×６列のマトリクス状に配列された容器群に、各列毎に異なる液を分注する。その後、各容器の外部に個々に設けられている磁力手段によって磁場を容器内部に作用させて、磁性体粒子と結合した結合体を各容器の内面に吸着保持させる。その後、前記ピペット手段１によって、内面に吸着保持された結合体を除く収容液を吸引して、排出用の別容器に移送して吐出することによって排除した後、磁力手段を容器から離間させることによって、又は電磁石をオフすることによって磁場を除去する。
【００８４】
ステップＳ２４で、図９に示すように前記移動手段３は、該反応用プレート１８₁ が載置されている回転ステージ２０₁ を９０°回転させることによって、マトリクス状に配列された容器群の行と列とを転置させる。
【００８５】
ステップＳ２５で、前記分注チップを前記洗浄槽１６で洗浄した後、又は分注チップを前記チップホルダーで交換した後、異分注試薬用容器１２から目的物質Ｂ₁ ，Ｂ₂ ，Ｂ₃ を別々に含む各試薬を各々１の量吸引して１／３の量ずつ又は１／３の量ずつ吸引して、行と列とを入れ換えた転置マトリクス状容器群に対して、ノズル群１０を並進移動することによって、既に各ウェルに収容された磁性体粒子と結合したＡ₁ 〜Ａ₃ の各結合体が収容されている容器に順次分注する。
【００８６】
ステップＳ２６で、反応及びインキュベーションにより、この段階で、転置マトリクスの第１列目の３個の容器には、各々磁性体粒子−Ａ₁ −Ｂ₁ 、磁性体粒子−Ａ₁ −Ｂ₂ 、磁性体粒子−Ａ₁ −Ｂ₃ という結合体が収容され、転置マトリクスの第２列目の３個の容器には、各々磁性体粒子−Ａ₂ −Ｂ₁ 、磁性体粒子−Ａ₂ −Ｂ₂ 、磁性体粒子−Ａ₂ −Ｂ₃ という結合体を含む液が収容され、第３列目の３個の容器には、各々磁性体粒子−Ａ₃ −Ｂ₁ 、磁性体粒子−Ａ₃ −Ｂ₂ 、磁性体粒子−Ａ₃ −Ｂ₃ の全９種類の結合体を含む液が各々１種ずつ収容されていることになる。
【００８７】
ステップＳ２７で、前記ノズル群１０は、並進移動することによって、前記転置マトリクスの容器群から順次、３個ずつ前記結合体が含まれた全収容液を容器から吸引して、前記液の１／３ずつ、即ち、最初の量の１／９ずつ転置されたマトリクス状容器の６列×３行及び３列×３行の全容器群に、前記９種類の結合体に各列毎に異なる結合体を含む液が分注され、各列に属する３個の各容器には同じ結合体を含む液が分注されるように、並進移動しながら分注する。その後、各容器の外部に個々に設けられている磁力手段によって容器内部に作用させて、磁性体粒子と結合した結合体を各容器の内面に吸着保持させ、前記ピペット手段１によって、内面に吸着保持された結合体を除く収容液を吸引して、排出用の別容器に移送して吐出することによって排除した後、磁力手段による磁場の作用を除去する。
【００８８】
ステップＳ２８で、前記移動手段３によって、図９に示すように、前記反応用プレート１８₁ 及び１８₂ を載置している各回転ステージ２０₁ 、２０₂ を９０°逆回転することによって、転置されたマトリクスの行と列を元の状態のマトリクスの行と列に戻す。
【００８９】
ステップＳ２９で、前記分注チップを前記洗浄槽１６で洗浄した後、又は分注チップを前記チップホルダーで交換した後、異分注試薬用容器１２から目的物質Ｃ₁ ，Ｃ₂ ，Ｃ₃ を別々に含む各試薬から各々１の量を吸引してその１／９の量ずつ又は１／９の量ずつ吸引して、３列×６行マトリクスの容器群及び３列×３行のマトリクスの容器群に対して、ノズル群１０を並進移動することによって、既に各ウェルに収容された磁性体粒子との結合体に、順次分注させる。
【００９０】
ステップＳ３０で、反応及びインキュベーションによって、マトリクスの第１列目の９個（６個と３個）の容器には、各々磁性体粒子−Ａ₁ −Ｂ₁ −Ｃ₁ 、磁性体粒子−Ａ₁ −Ｂ₂ −Ｃ₁ 、磁性体粒子−Ａ₁ −Ｂ₃ −Ｃ₁ 、磁性体粒子−Ａ₂ −Ｂ₁ −Ｃ₁ 、磁性体粒子−Ａ₂ −Ｂ₂ −Ｃ₁ 、磁性体粒子−Ａ₂ −Ｂ₃ −Ｃ₁ 、磁性体粒子−Ａ₃ −Ｂ₁ −Ｃ₁ 、磁性体粒子−Ａ₃ −Ｂ₂ −Ｃ₁ 、磁性体粒子−Ａ₃ −Ｂ₃ −Ｃ₁ の９種類の結合体が得られ、各々１種ずつ収容されていることになる。
【００９１】
マトリクスの第２列目の９個（６個と３個）の容器には、各々磁性体粒子−Ａ₁ −Ｂ₁ −Ｃ₂ 、磁性体粒子−Ａ₁ −Ｂ₂ −Ｃ₂ 、磁性体粒子−Ａ₁ −Ｂ₃ −Ｃ₂ 、磁性体粒子−Ａ₂ −Ｂ₁ −Ｃ₂ 、磁性体粒子−Ａ₂ −Ｂ₂ −Ｃ₂ 、磁性体粒子−Ａ₂ −Ｂ₃ −Ｃ₂ 、磁性体粒子−Ａ₃ −Ｂ₁ −Ｃ₂ 、磁性体粒子−Ａ₃ −Ｂ₂ −Ｃ₂ 、磁性体粒子−Ａ₃ −Ｂ₃ −Ｃ₂ の９種類の結合体が各々１種ずつ収容されていることになる。
【００９２】
マトリクスの第２列目の９個（６個と３個）の容器には、各々磁性体粒子−Ａ₁ −Ｂ₁ −Ｃ₃ 、磁性体粒子−Ａ₁ −Ｂ₂ −Ｃ₃ 、磁性体粒子−Ａ₁ −Ｂ₃ −Ｃ₃ 、磁性体粒子−Ａ₂ −Ｂ₁ −Ｃ₃ 、磁性体粒子−Ａ₂ −Ｂ₂ −Ｃ₃ 、磁性体粒子−Ａ₂ −Ｂ₃ −Ｃ₃ 、磁性体粒子−Ａ₃ −Ｂ₁ −Ｃ₃ 、磁性体粒子−Ａ₃ −Ｂ₂ −Ｃ₃ 、磁性体粒子−Ａ₃ −Ｂ₃ −Ｃ₃ の９種類の結合体が各々１種ずつ収容されていることになる。このようにして、全２７種類の結合体群が得られることになる。
【００９３】
ステップＳ３１で、該３列×６行、及び３列×３行のマトリクス状に配列されたウェルに解離剤を分注して、各々前記結合体を磁性体粒子から解離させ、磁性体粒子のみを分離することによって、最終生成化合物を得ることができる。
【００９４】
続いて、第四の実施の形態として、前記磁力手段が配設されたピペット手段による化学反応処理装置を用いて、構造解析を行うための標識化の処理について説明する。
図７に示すように、ステップＳ１で、前記入力手段６から、構造解析の処理過程の内容を特定するため、処理過程情報を入力する。処理過程を特定すると、構造解析を行う点、その試薬が収容された容器の位置、磁性体粒子が収容された容器の位置、反応が行われる容器の位置、ピペット手段又は処理ステージの移動の制御、反応時間、分注の順序等についても、予め前記記憶手段７に格納された情報に基づいて自動的に特定又は設定される。
【００９５】
ステップＳ２で、処理過程が構造解析なので、自動的にステップＳ５１に進む。ステップＳ５１で、前記ピペット手段のノズル群１０が前記処理ステージ２のチップホルダ１５に移動して３連分の分注チップを自動的に装着し、処理ステージ２の異分注試薬用容器１２にノズル群１０を移動させて、該容器１２に収容さているアミノ酸Ａ，Ｂ，Ｃを含む試薬を吸引して、図８に示すように、反応用プレート１８の６行×６列のマトリクス状に配置された容器群のうち３行×１列分の容器群に前記アミノ酸Ａ，Ｂ，Ｃを分注する。
【００９６】
ステップＳ５２で、該アミノ酸を識別するために、アミノ酸ＡにＴ₁ 、アミノ酸ＢにはＴ₂ 、アミノ酸ＣにはＴ₁ ＋Ｔ₂ というタグを対応させるために、別の異分注試薬用容器１２に貯溜されているこれらのタグを吸引して、前記容器に分注する。
【００９７】
ステップＳ５３で、該アミノ酸及びタグと反応により結合するアミノ基がコーティングされている磁性対粒子を前記アミノ酸及びタグが分注された容器に分注して混合する。その際、アミノ基の０．５〜１％にタグが結合し、残りのアミノ基に前記アミノ酸が結合するように、タグの分注量が設定されている。
【００９８】
したがって、この段階で、３個の各容器には、磁性体粒子−アミノ酸Ａ−タグＴ₁ 、磁性体粒子−アミノ酸Ｂ−タグＴ₂ 、磁性体粒子−アミノ酸Ｃ−タグＴ₁ ＋Ｔ₂ の各結合体が得られる。
【００９９】
ステップＳ５４で、前記磁性体粒子にアミノ酸及びタグが結合した各結合体を混合してランダム化する。
ステップＳ５５で、３個の容器に等量分割するとともにアミノ保護基を切断する。
【０１００】
ステップＳ５６で、遊離したアミノ酸に新しいタグＴ₃ ，Ｔ₄ ，Ｔ₃ ＋Ｔ₄ を３個の各容器に分注する。ここで、タグＴ₃ は、アミノ酸Ｘを識別するものであり、タグＴ₄ はアミノ酸Ｙを識別するものであり、タグＴ₃ ＋Ｔ₄ は、アミノ酸Ｚを識別するためのものである。
【０１０１】
ステップＳ５７で、アミノ酸Ｘ，Ｙ，Ｚを前記ピペット手段により前記３個の容器に各々分注する。
ステップＳ５８で、アミノ酸同士が反応及びインキュベーションにより結合する。この段階で、第一の容器には、Ｔ₁ Ｔ₃ −磁性体粒子−Ａ−Ｘ、Ｔ₂ Ｔ₃ −磁性体粒子−Ｂ−Ｘ、Ｔ₁ Ｔ₂ Ｔ₃ −磁性体粒子−Ｃ−Ｘ、第二の容器にはＴ₁ Ｔ₄ −磁性体粒子−Ａ−Ｙ、Ｔ₂ Ｔ₄ −磁性体粒子−Ｂ−Ｙ、Ｔ₁ Ｔ₂ Ｔ₄ −磁性体粒子−Ｃ−Ｙ、第三の容器にはＴ₁ Ｔ₃ Ｔ₄ −磁性体粒子−Ａ−Ｚ、Ｔ₂ Ｔ₃ Ｔ₄ −磁性体粒子−Ｂ−Ｚ、Ｔ₁ Ｔ₂ Ｔ₃ Ｔ₄ −磁性体粒子−Ｃ−Ｚが各々収容され、各アミノ酸の結合体が標識化されることになる。
【０１０２】
続いて、アッセイを行った後、ステップＳ５９で、取り出された磁性体粒子からタグを切断した後、ステップＳ６０でＥＣ−ＧＣによって分析を行うことによって、その磁性体粒子に結合しているアミノ酸配列を特定することができる。
【０１０３】
続いて、第五の実施の形態として、図１０及び図１１に基づいて、パラレル合成法(paralel synthesis) により、少なくとも磁力手段がピペット手段１の各ノズルに連通する液通路の外部に配設したものを用いて、３種のアミノ酸（Ａ，Ｂ，Ｃ）を含む試薬を使用して、トリペプチドを最終生成化合物として合成する他の例について説明する。
【０１０４】
図７のステップＳ１で、前記入力手段６から、コンビナトリアル合成の処理過程の内容を特定するため、処理過程情報を入力する。処理過程を特定すると、図７のステップＳ２で、パラレル合成を行う点、その試薬が収容された容器の位置、磁性体粒子が収容された容器の位置、反応が行われる容器の位置、ピペット手段又は処理ステージの移動の制御、反応時間、分注の順序等についても、予め前記記憶手段７に格納された情報に基づいて自動的に特定又は設定される。これによって、必要な試薬の配列等が前記制御手段４によって決定されることになる。
【０１０５】
該解析によって、本形態例では、３種類の物質群（Ａ，Ｂ，Ｃ）を用いて、成分の結合の数が３である最終生成物を生成するのであるから、第十二の発明の記号を用いると、ｎ＝３，ｍ＝９，ｋ＝３である。即ち、３列×９行容器群を３群必要とする。しかし、１つのプレートには、６列×６行のマトリクス容器群しか配列されていないので、各３列×６行容器群は、２つのプレートにまたがって配列されることになる。本形態例では次のようにして前記物質群を配置する。
【０１０６】
まず、前記ピペット手段１のノズル群１０が、前記処理ステージ２のチップホルダ１５に移動して６連の分注チップのうち３連分を自動的に装着し、処理ステージの異分注試薬容器１２にノズル群１０を移動させて、該容器１２に収容されているアミノ酸Ａ，Ｂ，Ｃを含む液を各々吸引して、図１０（ａ）の反応用プレート１８₁ ，１８₂ 及び図１１（ｄ）の反応用プレート１８₃ ，１８₄ の６行×６列のマトリクス状に配列された４つの容器群に各々前記アミノ酸Ａ，Ｂ，Ｃをマトリクス状に分注及び配置する。ここでは、前記ノズル群１０は１列×３行のベクトル状に配列されているものとする。
【０１０７】
また、各反応用プレート１８₁ ，１８₂ ，１８₃ ，１８₄ は各々回転ステージ２０₁ ，２０₂ ，２０₃ ，２０₄ に設けられている。反応用プレート１８₁ 上では、６列×６行の各列の容器群毎にＡ，Ｂ，Ｃの順で分注配置し、３列×６行の各マトリクスを各々容器群Ｍ₁ ，容器群Ｍ₂ と表す。反応用プレート１８₂ 上では、６列×３行の各列の容器群毎にＡ，Ｂ，Ｃの順で配置し、３列×３行の各マトリクスを各々容器群Ｍ₃ ，容器群Ｍ₄ と表す。また、図１１（ｄ）の反応用プレート１８₃ 上では、３列×６行の各列の容器群毎にＡ，Ｂ，Ｃの順で配置し、容器群Ｍ₅ と表す。図１１（ｄ）の反応用プレート１８₄ 上では、３列×３行の各列の容器群毎にＡ，Ｂ，Ｃの順で配置し、容器群Ｍ₆ と表す。即ち、容器群Ｍ₁ とＭ₃ とは第１群の３列×９行容器群に相当し、容器群Ｍ₂ とＭ₄ とは前記第２群の３列×９行容器群に相当し、容器群Ｍ₅ とＭ₆ とは前記第３群の３列×９行容器群に相当する。
【０１０８】
次に、第１群の３列×９行容器群に相当する前記反応用プレート１８₁ の３列×６行の容器群Ｍ₁ 及び反応用プレート１８₂ の３列×３行の容器群Ｍ₃ に対して、前記ピペット手段１のノズルによって、前記Ａ，Ｂ，Ｃと特異的に反応して結合する物質がコーティング又は保持された磁性体粒子が懸濁された液を等量ずつ分注して、磁性体粒子と前記Ａ，Ｂ，Ｃの各アミノ酸と結合させて結合体を生成させる。
【０１０９】
次に、磁性体粒子と各アミノ酸の結合体が生成されて収容されている前記第１群の３列×９行容器群に相当する容器群Ｍ₁ 及び容器群Ｍ₃ を移送元とし、該容器群から、結合体のみを移送対象として、第２群の３列×９行容器群に相当する反応用プレート１８₁ 、１８₂ の容器群Ｍ₂ 及びＭ₄ を移送先として、前記回転ステージ２０₁ 、２０₂ を転置させずに移送する。
【０１１０】
前記ピペット手段１の３連の各ノズルは、最初に容器群Ｍ₁ の第１列にある（Ｍ₁ ）₁₁，（Ｍ₁ ）₁₂，（Ｍ₁ ）₁₃の３個の容器（添字は、列及び行番号を表す）に並進移動される。移送元である該容器群から、収容されている全液を吸引し吐出する際に磁力手段により液通路に磁場を及ぼすことによって、結合体を分離し、移送先にある容器群Ｍ₂ の第１列にある（Ｍ₂ ）₁₁，（Ｍ₂ ）₁₂，（Ｍ₂ ）₁₃の３個の容器にピペット手段１を並進移動させる。続いて、該３個の容器（Ｍ₂ ）₁₁，（Ｍ₂ ）₁₂，（Ｍ₂ ）₁₃から、収容されている液を磁力手段による磁場を液通路に及ぼさずに吸引、吐出することによって結合体をピペット手段１から各容器に移す。
【０１１１】
同様にして、前記ピペット手段１のノズル群１０を、前記結合体が収容されている容器群Ｍ₁ のうち、第１列にある（Ｍ₁ ）₁₄，（Ｍ₁ ）₁₅，（Ｍ₁ ）₁₆の３個の容器に並進移動する。移送元である該容器群から、収容されている全液を吸引し吐出する際に磁力手段により液通路に磁場を及ぼすことによって、結合体を分離し、移送先にある容器群Ｍ₂ の第２列にある（Ｍ₂ ）₂₁，（Ｍ₂ ）₂₂，（Ｍ₂ ）₂₃の３個の容器にピペット手段１を並進移動させる。続いて、該３個の容器（Ｍ₂ ）₂₁，（Ｍ₂ ）₂₂，（Ｍ₂ ）₂₃から、収容されている液を磁力手段による磁場を液通路に及ぼさずに吸引、吐出することによって結合体をピペット手段１から各容器に移す。
次に、前記ピペット手段１のノズル群１０を、前記結合体が収容されている容器群Ｍ₃ のうち、第１列にある（Ｍ₁ ）₁₄，（Ｍ₁ ）₁₅，（Ｍ₁ ）₁₆の３個の容器に並進移動する。移送元である該容器群から、同様にして結合体を分離し、移送先にある容器群Ｍ₂ の第３列にある（Ｍ₂ ）₃₁，（Ｍ₂ ）₃₂，（Ｍ₂ ）₃₃の３個の容器にピペット手段１のノズル群１０を並進移動させ、同様にして該容器群に、結合体をピペット手段１から各容器に移す。
【０１１２】
以下同様にして、３個の容器群をまとめて１つの要素とする仮想的マトリクス上、相互に転置位置となるように、Ｍ₁ 及びＭ₃ の第１群の３列×９行容器群から、Ｍ₂ 及びＭ₄ の第２群の３列×９行容器群に結合体のみを移送すると、図１０（ｂ）に示すように結合体及び試薬が容器群Ｍ₂ 及びＭ₄ の第２群の３列×９行容器群に配置される。
即ち、この段階で反応及びインキュベーションによって、磁性体粒子−Ａ−Ａ，磁性体粒子−Ａ−Ｂ，磁性体粒子−Ａ−Ｃ，磁性体粒子−Ｂ−Ａ，磁性体粒子−Ｂ−Ｂ，磁性体粒子−Ｂ−Ｃ，磁性体粒子−Ｃ−Ａ，磁性体粒子−Ｃ−Ｂ，磁性体粒子−Ｃ−Ｃが生成されることになる。
【０１１３】
続いて、反応用プレート１８₁ 及び１８₂ が設けられている回転ステージ２０₁ 及び２０₂ を９０°回転して転置することによって、図１１（ｃ）に示すように、反応用プレート１８₁ 及び反応用プレート１８₂ に設けられた第２の３列×９行容器群Ｍ₂ 及びＭ₄ は転置されて、各々容器群Ｍ₂ ^T 及びＭ₄ ^T （“Ｔ”は転置行列を表す）と配列されることになる。
【０１１４】
次に、前記ピペット手段１のノズル群１０は、例えば、容器群Ｍ₂ の第６列にある３個の容器（Ｍ₂ ^T ）₆₁，（Ｍ₂ ^T ）₆₂，（Ｍ₂ ^T ）₆₃の収容物を全量吸引する際に外部に設けた磁力手段によりピペット手段１の液通路に磁場を及ぼし、前記結合体をピペット手段の液通路内壁に吸着保持することによって分離する。結合体を分離すると、移送先の第３群の３列×９行容器群である反応用プレート１８₃ にある容器群Ｍ₅ の第１列にある（Ｍ₅ ）₁₁，（Ｍ₅ ）₁₂，（Ｍ₅ ）₁₃の３個の容器に移動させる。続いて、該（Ｍ₅ ）₁₁，（Ｍ₅ ）₁₂，（Ｍ₅ ）₁₃の３個の容器から、収容されている液を、磁力手段による磁場を液通路に及ぼさずに吸引、吐出することによって結合体をピペット手段１から各容器に移す。
【０１１５】
同様にして、ピペット手段１のノズル群１０を、前記結合体が収容されている容器群Ｍ₂ ^T の第５列にある（Ｍ₂ ^T ）₅₁，（Ｍ₂ ^T ）₅₂，（Ｍ₂ ^T ）₅₃に並進移動させる。移送元である該容器群の収容物を全量吸引する際に磁力手段をピペット手段１の液通路に磁場を及ぼし、前記結合体を分離する。結合体を分離すると、移送先の反応用プレート１８₃ の容器群Ｍ₅ の第２列にある（Ｍ₅ ）₂₁，（Ｍ₅ ）₂₂，（Ｍ₅ ）₂₃の３個の容器に移動させる。続いて、該（Ｍ₅ ）₂₁，（Ｍ₅ ）₂₂，（Ｍ₅ ）₂₃の３個の容器から、収容されている液を、磁力手段による磁場を液通路に及ぼさずに吸引、吐出することによってピペット手段が保持していた結合体を各容器に移す。
【０１１６】
同様にして、順次Ｍ₂ ^T の容器群から、結合体のみを分離して、容器群Ｍ₅ の対応する各容器群に移送し、最終的には（Ｍ₄ ^T ）₁₁，（Ｍ₄ ^T ）₁₂，（Ｍ₄ ^T ）₁₃を反応用プレート１８₄ にある容器群Ｍ₆ の第３列にある（Ｍ₆ ）₃₁，（Ｍ₆ ）₃₂，（Ｍ₆ ）₃₃に移送することになる。
【０１１７】
こうして最終生成物として、磁性体粒子−Ａ−Ａ−Ａ，磁性体粒子−Ａ−Ａ−Ｂ，磁性体粒子−Ａ−Ａ−Ｃ，磁性体粒子−Ａ−Ｂ−Ａ，磁性体粒子−Ａ−Ｂ−Ｂ，磁性体粒子−Ａ−Ｂ−Ｃ，磁性体粒子−Ａ−Ｃ−Ａ，磁性体粒子−Ａ−Ｃ−Ｂ，磁性体粒子−Ａ−Ｃ−Ｃ，磁性体粒子−Ｂ−Ａ−Ａ，磁性体粒子−Ｂ−Ａ−Ｂ，磁性体粒子−Ｂ−Ａ−Ｃ，磁性体粒子−Ｂ−Ｂ−Ａ，磁性体粒子−Ｂ−Ｂ−Ｂ，磁性体粒子−Ｂ−Ｂ−Ｃ，磁性体粒子−Ｂ−Ｃ−Ａ，磁性体粒子−Ｂ−Ｃ−Ｂ，磁性体粒子−Ｂ−Ｃ−Ｃ，磁性体粒子−Ｃ−Ａ−Ａ，磁性体粒子−Ｃ−Ａ−Ｂ，磁性体粒子−Ｃ−Ａ−Ｃ，磁性体粒子−Ｃ−Ｂ−Ａ，磁性体粒子−Ｃ−Ｂ−Ｂ，磁性体粒子−Ｃ−Ｂ−Ｃ，磁性体粒子−Ｃ−Ｃ−Ａ，磁性体粒子−Ｃ−Ｃ−Ｂ，磁性体粒子−Ｃ−Ｃ−Ｃが、図１１の（ｄ）の第３群の３列×９行容器群の２７個の各容器に収容されることになる。
【０１１８】
以上説明した処理ステージ２では、各プレート１８毎に回転ステージ２０が設けられ、プレート１８毎に回転可能であったが、該場合に限られるものではなく、例えば、図１２に示す処理ステージ３５では、複数のプレート１８（この例では４個のプレート）が１つの回転ステージ３６上に載置され、複数のプレートが同時に回転できるように設けても良い。
以上説明した実施の形態では、スプリット合成法及びパラレル合成法についてのみ説明したが、両者を組み合わせた合成法であっても良い。
さらに、本発明に係る合成法は、生体高分子のみならず有機物及び無機物に適用することが可能である。
【０１１９】
前述したスプリット合成法では、混合を２回行い、混合液のａ（＝３）個の等分割を２回行うようにしたが、本発明はこのような場合に限られず、ａ個の等分割をｎ回繰り返すことによって、ａⁿ 個の目的化合物群を得ることができる。
また、該スプリット合成法は、トリペプチドをライブラリとして合成する例を示したが、該例に限られず、例えば、他の蛋白質、ＤＮＡや多糖糖の生体高分子のみならず、他の有機物や無機物糖にも本発明を適用することができる。
【０１２０】
以上の例では、測定手段について説明しなかったが、測定手段を設け、さらに測定結果の解析をも行うような装置を設けることによって、測定から解析までを一貫して行うことができる。
【０１２１】
また、１種類の磁性体粒子を用いる場合のみならず、２以上の種類の磁性体粒子の中から、必要な種類の磁性体粒子を処理過程の内容に基づいて選択して使用するようにしても良い。これによって、反応に関与する物質と特異的に反応する物質を磁性体粒子にコーティングしたり、反応に関与する物質に悪影響を与えない物質を選択してコーティングしたりすることによって、適用することができる処理の可能性の幅が増大し汎用性があり、且つ信頼性のある化学反応処理を行うことが出来る。
さらに、前記各実施の形態に用いた物質群の例は、説明の便宜のために、簡単な例を挙げて説明したものに過ぎず、化学反応処理の内容、目的物質の種類、数、最終生成物等は、これらの例に限定されるものではない。
【０１２２】
【発明の効果】
以上説明したように、第一の発明では、磁力手段が配設されたピペット手段又は処理ステージの一部若しくは全体を転置移動又は並進移動可能を可能とする移動手段を設けるとともに、指定手段による指定に基づいて、試薬等の物質群の配置、各容器の収容物のうち移送対象、移送元及び移送先の容器若しくは容器群、移送の際の転置移動若しくは並進移動の別、並びに、ピペット手段の動作を特定して指示する制御手段を設けている。
【０１２３】
したがって、多数の物質群をマトリクス状に配置し、転置移動及び並進移動を行うことにより、単純で無駄のない動作で効率良く、迅速にシステマチックに化学反応処理を行うことができるとともに、装置の構成を簡単でコンパクトにすることができる。
【０１２４】
また、磁力手段を配設することによって、各容器内の収容物の中から移送対象を選択特定することによって、処理を簡略化又は多様化することができるとともに、信頼性良く、種々の処理をコンパクトで簡単な装置で行うことができる。
多数で少量の物資群を扱うことによって、最終生成化合物の多品種少量生産を効率良く行うことができる。
【０１２５】
第二の発明では、処理過程の内容等に応じて、ピペット手段のマルチノズルの個数や非配列、処理ステージの容器群の個数や配列を設定することによって、無駄のない効率的な利用と、全体の装置をコンパクトにすることができる。
【０１２６】
第三の発明は、ピペット手段の分注器のノズルに着脱可能に装着する分注チップを用いることによって、洗浄を必要とせずに、クロスコンタミネーションのない信頼性のある化学反応処理を行うことができる。
第四の発明は、容器群の一部又は全体に、外部に磁力手段を設けて各容器内面での磁性体粒子の吸着脱着が可能としたものである。
これによって、容器に収容された収容物の中から磁性体粒子との結合体又は結合体以外の物を選択して移送することができるので、多様な処理が可能となる。
【０１２７】
第五の発明は、容器に蓋部材を設けることによって、容器内の液体の蒸発、汚染又は発生する気体を閉じ込めて環境への悪影響を防止するとともに、ピペット手段の外側への付着物を拭うことができるので扱いやすい。
【０１２８】
第六の発明は、前記処理ステージに、処理に必要な種々の装置を設けることによって、化学反応処理を一貫して又は効率良く行うことができる。
第七の発明は、処理ステージ等を密閉空間内に設けることによって、人体等に有害な化学反応を封じ込めて環境を保護したり、密閉空間内に種々雰囲気、条件又は状況を現出することができるので、多様性のある処理を行うことができる。
【０１２９】
第八の発明は、処理時間等を小さくすることによって、化学反応処理を効率良く、又は迅速に行うことができる。
第九の発明は、磁力手段が配設されたピペット手段、処理ステージ、移動手段及び制御手段を処理過程の内容に基づいて制御を行うように用いている。
したがって、第一の発明で奏する効果と同様に、処理過程の内容に基づいて容易に、効率良く又は迅速に化学反応処理を行うことができるとともに、装置の構成を簡単でコンパクトにすることができる。
【０１３０】
第十の発明によれば、簡単な構造と動作によって、いわゆるスプリット合成法等の処理を効率的、迅速に行うことができる。
第十一の発明又は第十二の発明によれば、各容器の収容物のうち磁性体粒子との結合体を分離して、結合体以外の収容物を除去しながら、物質群との間で反応を行うようにしているので、目的物質以外の不純物や老廃物を除去して信頼性のある処理を効率的、迅速に行うことができる。
【０１３１】
第十三の発明は、識別子を磁性体粒子に関連付けることによって、磁性体粒子したがって、該磁性体粒子に結合する物質を標識化することができる。
これによって、合成等がされた目的物の構造解析を容易且つ確実に行うことができる。
【図面の簡単な説明】
【図１】本発明の実施の形態に係るブロック図
【図２】本発明の実施の形態に係る処理ステージを示す平面図
【図３】本発明の実施の形態に係る同一試薬分注用容器の第一の例を示す斜視図
【図４】本発明の実施の形態に係る同一試薬分注用容器の第二の例を示す斜視図
【図５】本発明の実施の形態に係る同一試薬分注用容器の第三の例を示す図
【図６】本発明の実施の形態に係る洗浄槽及びチップホルダを示す図
【図７】本発明の実施の形態に係る流れ図
【図８】本発明の実施の形態に係るスプリット合成法の説明図
【図９】本発明の実施の形態に係るパラレル合成法の説明図
【図１０】本発明の実施の形態に係る他のパラレル合成法の説明図（１）
【図１１】本発明の実施の形態に係る他のパラレル合成法の説明図（２）
【図１２】本発明の実施の形態に係る他の処理ステージを示す平面図
【符号の説明】
１…ピペット手段
２，３５…処理ステージ
３…移動手段
４…制御手段
５…表示手段
６…入力手段
７…記憶手段
１０…ノズル群
１１、１３…同一分注試薬用容器
１２、１４…異分注試薬用容器
１６…試薬分注ノズル洗浄槽
１７…反応ノズル洗浄槽
１８…反応用プレート
１９…混合用プレート
２０…回転ステージ[0001]
BACKGROUND OF THE INVENTION
The present invention relates to combinatorial chemistry for efficiently synthesizing a large number of compound groups (libraries) using combinations and utilizing those compounds in accordance with various purposes. The present invention relates to a chemical reaction processing apparatus and method using magnetic particles used for synthesizing at once, performing an activity test, and performing various processes such as structure determination.
[0002]
[Prior art]
Conventionally, the reaction field where combinatorial synthesis is performed is currently performed in either solid phase or liquid phase, but in the case of combinatorial synthesis, the solid phase method is more than the liquid phase method considering the speed and ease of automation. The solid-phase method is now the mainstream.
[0003]
However, solid-phase synthesis using non-magnetic beads (resin) is more difficult to control the reaction than liquid-phase synthesis, and byproducts cannot be removed. It had the problem that.
[0004]
In addition, even in the case of solid-phase synthesis using non-magnetic beads, automatic loading and unloading of containers are very difficult, and the supernatant and precipitate fractions can be mechanically performed. It was very difficult, and it was difficult to perform full processing completely, including reagent addition, stirring, temperature control, dialysis, filtration, etc. .
[0005]
[Problems to be solved by the invention]
The present invention was devised in view of the current situation. The object of the present invention is to achieve a series of chemical reaction treatments automatically and consistently by using magnetic particles. It is an object of the present invention to provide a chemical reaction processing apparatus and method using magnetic particles that can be used.
[0006]
Secondly, the first principle and the "the method of controlling the desorption of a magnetic material using a dispenser and various devices processed by this method" previously proposed by the present applicant (Japanese Patent Application No. 7-39425). It is an object of the present invention to provide a chemical reaction processing apparatus and method using magnetic particles capable of performing efficient chemical reaction processing by combining with the principle of (1).
[0007]
Third, by controlling the magnetic particles, including operations that have the same meaning as the processing such as addition, stirring, temperature control, extraction, solvent evaporation, filtration, concentration, separation, dialysis, etc. An object of the present invention is to provide a chemical reaction processing apparatus and method using magnetic particles that can reduce the scale of the apparatus and reduce the cost because the chemical processing can be performed consistently.
[0008]
Fourth, by confining a device with a reduced size in a sealed space, it prevents the diffusion of substances harmful to the human body and does not adversely affect the environment, or isolates the air and treats it in various atmospheres. It is an object of the present invention to provide a chemical reaction treatment apparatus and method using magnetic particles capable of performing the above.
[0009]
Fifth, magnetic particles that are more efficient, more reliable, and more versatile and more versatile than solid phase methods using non-magnetic particles can be processed. It is an object of the present invention to provide a used chemical reaction processing apparatus and method.
[0010]
Sixth, by arranging and arranging a large number of container groups and substance groups in a matrix, performing translocation movement or translational movement, and specifying the objects to be transferred in each container, it is possible to operate simply and without waste. It is an object of the present invention to provide a chemical reaction processing apparatus and method using magnetic particles that can perform chemical reaction processing efficiently, quickly and systematically.
Seventh, it is possible to deal with a large variety of small-quantity production by efficiently handling a small but many kinds of substance groups.
[0011]
[Means for Solving the Problems]
In order to solve the above problems, a first invention is a pipette means having a multi-nozzle capable of sucking and discharging liquid, a processing stage provided with a container group in which containers are arranged in a matrix, and the pipette Magnetic means provided in a part or the whole of the container group of the means or the processing stage and capable of adsorbing and desorbing magnetic particles on the liquid passage of the pipette means or the inner surface of each container, and the nozzle group of the pipette means or the processing Contents of chemical reaction processing such as combinatorial synthesis, moving means that enables transposition movement that moves by exchanging rows and columns and translation movement that moves without changing rows and columns, for a part or the whole of the container group of the stage And a control means for controlling the operation of each means, and the control means controls the magnetic particles based on the designation by the designation means. A group of substances on the container group, objects to be transferred among the contents accommodated in each container, containers or container groups of a transfer source and a transfer destination, apart from transposition movement or translation movement at the time of transfer, In addition, operations such as suction of the pipette means and adsorption by the magnetic force means are specified and each means is instructed.
[0012]
Here, the chemical reaction process includes, for example, various processes such as an activity test and a structure determination in addition to the combinatorial synthesis process.
As the “magnetic means”, for example, an electromagnet provided close to the outside of the liquid passage of the pipette means or the container or container group of the processing stage, or provided so as to be able to approach and separate from the outside of the liquid passage, container or container group. There are cases of permanent magnets.
The magnetic means may be provided on both the pipette means or the processing stage.
“Contents of treatment process” includes various contents related to treatment, for example, the substance group involved in the reaction, the type, number, amount, mixing, reaction, order, time, temperature condition, etc. of the target substance group. .
In addition, the types of target substance groups include those having different concentrations by providing a concentration gradient for the same substance.
[0013]
The “container” includes holes and wells provided in the plate.
The arrangement of “pipette means having multi-nozzles” includes not only a case where they are arranged in one column (row) but also a case where they are arranged in a matrix.
[0014]
By the pipette means and the magnetic force means, operations such as separation, agitation, and washing can be efficiently performed as well as dispensing by simply sucking or discharging a liquid.
Here, in the case where the magnetic force means is disposed in the pipette means, for example, a pipette having a separation region portion in which a magnetic field action is exerted in a liquid passage connecting the tip portion and the reservoir portion, and sucking or discharging the liquid. When the liquid in which the magnetic particles are suspended is passed through the separation region, the magnetic field acts on the separation region from the side surface of the liquid passage, and the magnetic particles are adsorbed on the inner surface of the liquid passage. Separation is performed by
[0015]
In addition, when the magnetic means is provided on the processing stage, not only when the magnetic means is provided for each container, but also to apply a magnetic field to the entire container group, for example, a strong magnetic force common to the bottom surface of the container group. A magnetic field may be applied to each container by providing means.
The “object to be transferred among stored items stored in each container” may include a part or the whole of the stored items. Moreover, as a part of the container, only the combined body with the magnetic particles, the stored object excluding the combined body, or a quantitative part of the stored object may be used.
[0016]
Here, the “matrix shape” means a state in which a plurality of elements are arranged or arranged along at least two directions on the plane (row direction and column direction) or in parallel. An arrangement or arrangement along the row direction is called a row, and an arrangement or arrangement along the column direction is called a column. The row direction and the column direction do not necessarily need to intersect at 90 °, and include cases where they are arranged obliquely. Of the matrix arrangement or arrangement, the matrix shape of 1 row × multiple columns (1 row and 1 column) or multiple rows × 1 column is particularly referred to as “vector”.
The “matrix arrangement” includes a case where a plurality of container groups arranged in a matrix are further arranged in a matrix.
[0017]
“Transposition movement” is to move by moving rows and columns arranged in a vector or matrix, and when the rows and columns are arranged at an angle of 90 °. , 90 ° rotational movement. “Translational movement” refers to movement without replacement. In order to be transposable, it is necessary to match both the row and column intervals of the processing stage matrix and the nozzles of the pipette means.
As the “translocation movement”, for example, the pipette means may be rotated by 45 °, the container group may be rotated by −45 °, and the total may be rotated by 90 °.
[0018]
An appropriate type of “magnetic particles” is selected in order to bind related substance groups according to the content of the treatment process. For example, when a magnetic substance particle is made to hold a participating substance in a porous material provided with a large number of recesses, or by adsorbing the participating substance or coating or holding a substance that reacts specifically with the participating substance. , Which binds the participating substances. At that time, the substance involved by coating or holding a magnetic substance particle with a linker having a functional group for binding and cleaving the substrate and the carrier used in the connecting portion of the low molecular weight compound such as an amino acid serving as the substrate and the insoluble carrier. You may make it hold | maintain a group.
As the magnetic particles, various particles may be selected and used according to the content of the treatment process in order to bind the substances involved.
[0019]
The predetermined container of the “processing stage” may be provided with accessories such as a low-temperature or high-temperature thermostatic device or a measuring device.
“Arrangement of substance groups on the container group” refers to how each substance group is arranged in each container group provided in the process stage in the process of handling a large number of substance groups such as combinatorial synthesis. The positioning of the substance group is very important in order to improve the processing efficiency and identify each substance group. Including such an arrangement, the control means not only moves the pipette means but also organically links the movement of the processing stage container group, the arrangement of the substance group, etc. so as to be efficient or quick. To control.
[0020]
For example, if each processing procedure is described as an operation similar to a vector operation or a matrix operation by arranging the arrangement of the nozzle group and the container group of the pipette means into a vector shape or a matrix shape, many kinds of substance groups can be represented. Control of the processing handled becomes efficient.
If a simple example with four types of reagents is described, the reagent a_i(I = 1, 2) is stored in a 2 × 1 container group, and reagent b_jWhen (j = 1, 2) is also stored in a 2 × 1 container group, each reagent is reacted to a_ib_j(A)₁, A₂)^T(B₁, B₂) From the product of the matrix (vector)₁b₁, A₁b₂, A₂b₁, A₂b₂And the following matrix is obtained ("T" represents a transposed matrix).
[Expression 1]

This formula includes information on how to arrange or move the reagents (including transposition) and what kind of target substance is obtained in which container.
[0021]
According to a second aspect of the present invention, in the first aspect, the arrangement of the nozzles of the pipette means or the arrangement of the containers of the processing stage is related to each other and determined based on the contents of the processing process. .
[0022]
Here, the arrangement of the nozzles includes not only the case of one column (row) but also the case of arrangement in a matrix. As an example of “related to each other”, each diameter is determined so that the nozzle can enter without contacting each container, and when the distance between the nozzles and the distance between the container groups coincide, the distance between the nozzles is This includes the case where the interval between the groups is an integral multiple, or the number of nozzle groups arranged and the number of rows or columns of the container group are matched or made an integral multiple or a fraction thereof. In addition, as an example of “contents of treatment process”, containers arranged in a matrix having the number of rows equal to the number of types of substance groups necessary for the reaction or the number of columns of different substance group types Groups may be used.
[0023]
According to a third invention, in the first invention or the second invention, the pipette means has a dispensing tip that is detachably attached to a nozzle of a dispenser of a liquid suction line.
Here, for example, the dispensing tip is formed of plastics, stainless steel or Teflon.
[0024]
According to a fourth aspect of the present invention, in the first to third aspects of the present invention, a part of the container group of the processing stage or a part of the whole container is provided with a magnetic means on the outside, and the magnetic particles on the inner surface of each container Can be adsorbed and desorbed.
Thereby, the said control means is controllable so that a thing other than a conjugate | bonded_body or its part may be transferred as a transfer object among the things accommodated in each container.
[0025]
According to a fifth invention, in the first invention, a part or a whole of the container group provided on the processing stage is covered with a film-like or plate-like lid member and attached to the nozzle of the pipette means. It is possible to penetrate through the tip of the dispensed tip.
In order to be “penetrable”, there are a case where a penetrating hole is previously provided in the lid member and a case where a lid member not previously provided with a hole is penetrated at the tip of the dispensing tip. Examples of such a lid member include a laminate and a rubber plate.
[0026]
6th invention is 1st invention thru | or 5th invention, Processing process of the container for dispensing, the container for washing | cleaning, the container for reaction, a thermostat or / and the apparatus for a measurement in the said process stage Necessary equipment is provided.
[0027]
According to a seventh invention, in any one of the first to sixth inventions, the processing stage and the pipette means are provided in an airtight container.
In an eighth aspect based on the first aspect to the seventh aspect, the control means is a processing time during a processing process with respect to the pipette means, the processing stage or the moving means, or the moving means. Control is efficiently performed by reducing the movement amount, the movement time, or the energy consumption, or by simplifying the operation.
[0028]
The ninth invention is arranged based on the contents of the designated processing steps by using the pipette means, the processing stage, the designation means, the moving means and the control means according to the first to eighth inventions. The process (S1) of arranging a plurality of types of substance groups such as reagents in a part or the whole of the container group of the processing stage and the substance group arranged based on the contents of the designated processing process A process of dispensing a liquid containing magnetic particles in a part or the whole of the container group, and combining the target substance in each substance and the magnetic particles by reaction or the like (S2), and designation Based on the contents of the processed process, a transposition movement in which a part or the whole of the nozzle group of the pipette means or the container group of the processing stage moves by exchanging rows and columns or a translation movement that moves without exchanging the rows and columns. By Some or all of the original container or containers stowed contained object in the group, is intended to include the steps (S3) for transporting the transfer destination of the container or container unit.
[0029]
Here, in order to transfer only the combined body of the contents, the magnetic means provided in the pipette means applies a magnetic field to the liquid passage of the pipette means to separate the magnetic particles and hold them on the inner wall. Do.
When something other than the combined body is transferred, for example, a magnetic body is provided in the container so that the combined body is adsorbed and held on the inner wall of the container.
[0030]
In a tenth aspect based on the ninth aspect, the step (S1) includes: a among a group of containers arranged in a matrix.₁~ A_nDispense and arrange equal amounts of reagents in n columns x 1 row of containers, b₁~ B_mIs a process of dispensing and arranging equal amounts of each reagent in a container group of m columns × 1 row, wherein the step (S2) dispenses magnetic particles into the container group in which the reagent is disposed, and a₁~ A_nIn this process, the target substance and the magnetic particles in the reagent are combined by reaction or the like to generate a combined body, and the step (S3) includes pipetting means for the entire contents in each container containing the combined body. The mixture is transferred into a common mixing container and mixed, and the mixed solution is divided into m equal parts, and an equal amount is pipetted into a container group in each column and m rows of containers arranged in a matrix of n columns and m rows. And select translation or transposition during the transfer, b₁~ B_mThis reagent is divided into n equal parts and transferred by pipette means to the container group in each column and n rows of containers arranged in a matrix of n columns × m rows.
Here, in the case of n = m, m kinds of reagents can be dispensed with a minimum number of times without omission, so that there is no need for transposition movement. Whether the displacement movement or the translation movement depends on the arrangement of nozzle groups of the pipette means.
In the present invention, chemical reaction treatment is performed by a so-called split pool method.
[0031]
In an eleventh aspect based on the ninth aspect, the step (S1) includes: a among a group of containers arranged in a matrix.₁~ A_nDispense and arrange equal amounts of each reagent in an n column x m row container group, b₁~ B_mIs a process of dispensing and arranging equal amounts of each reagent in a group of containers of m columns × 1 row, wherein the step (S2) is performed in an equal amount in all containers arranged in a matrix of n columns × m rows. Dispensing the magnetic particles to bind the target substance and the magnetic particles in the reagent, and the step (S3) is performed on all containers arranged in a matrix of the n columns × m rows, A magnetic field is applied by a magnetic means provided outside the container, and a combined body with magnetic particles is adsorbed and held on the inner surface of each container, and accommodation other than the combined body adsorbed and held on the inner surface of the container by the pipette means. Objects are removed by suction, and all the n columns × m rows arranged in a matrix of the n columns × m rows in which the combined bodies are accommodated are transferred by moving the container groups of the n columns × m rows or the nozzle groups of the pipette means. B.₁~ B_mIn this process, the reagent contained in each container is divided into n equal parts and transferred.
The present invention shows a parallel synthesis method that is performed when the magnetic means is disposed on the container side of the container of the processing stage.
[0032]
In a twelfth aspect based on the ninth aspect, the step (S1) includes the steps of: a / n groups of n columns × m rows arranged in a matrix of n columns × m rows,₁~ A_nThe n reagents of each type are dispensed in equal amounts into m groups of containers in each row, and the step (S2) is performed in each of the n rows of m / n groups × m. In the row container group, an equal amount of magnetic particles are dispensed to all the containers of the first group of n columns × m row container groups, and the target substance and the magnetic particles contained in each reagent are combined. The step (S3) is a step of generating a combined body using a pipette means in which only the combined body is provided with magnetic means out of the contents of each container of each n column × m row container group. Step of sequentially transferring from one group of n columns × m rows of container groups to the next group of n columns × m rows of containers by transposition or translation until reaching the m / nth group of n columns × m rows of containers It is.
[0033]
The present invention corresponds to a parallel synthesis method in which a magnetic means is provided in a pipette means.
“M / n (= k)” is component a contained in the final product_i(Where the target substance contained in each reagent is represented) is the number of bonds, and the final product is a_i1-A_i2-...- a_ik(I1, i2,..., Ik = 1, 2, ... n).
For example, if the end product is a_i-A_j-A_kIn the case of such a three-component system, three n column × m row container groups are prepared in advance in the processing stage.
[0034]
According to a thirteenth invention, in any of the ninth to twelfth inventions, an identifier for identifying each substance is added together with each substance of the substance group, and the identifier is added to the magnetic particles. The substance bonded to the magnetic particles is identified by being positioned around the bond or the magnetic particles.
Here, the “identifier” includes not only a tag but also an information storage chip.
“Located in the vicinity” includes the case of being trapped in a permeable capsule.
[0035]
DETAILED DESCRIPTION OF THE INVENTION
Based on FIGS. 1-6, 1st embodiment of this invention is described.
FIG. 1 shows a case where a chemical treatment apparatus using magnetic particles according to the present embodiment is applied to combinatorial synthesis treatment. As shown in the figure, the apparatus according to the present embodiment includes a pipette means 1 having a multi-nozzle for sucking and discharging liquids, and a group of containers for storing liquids arranged in a matrix along the xy axis. A processing stage 2 having at least a plate, and a magnetic passage (not shown) capable of adsorbing and desorbing magnetic particles on a liquid passage communicating with each nozzle of the pipette means 1 or an inner surface of each container of each processing stage 2. , Translational movement that moves without changing the rows and columns of the nozzle group and the container group along the xy direction and z direction along the board surface of the processing stage 2 and the transposition that moves by changing the rows and columns. And a moving means 3 comprising a rotatable motor or the like for moving the translating stage of the processing stage 2 so that a part of the container group is transposed in the plane. That.
[0036]
Further, the present chemical treatment apparatus includes an input means 6 corresponding to a designation means for designating the contents of chemical reaction processing such as combinatorial synthesis, suction and discharge operations of the pipette means 1, and magnetic particles using the magnetic force means. Adsorption, detachment, etc., arrangement of participating substance groups with respect to the container group of the processing stage, transfer object among the contents accommodated in each container, and a transfer source and transfer destination container or container group, transfer In accordance with the contents of the processing process designated by the input means 6 or the like, the transposition movement in which the row and the column are exchanged or the translation movement in which the row and the column are not exchanged are identified. Control means 4 comprising a CPU or the like for instructing each means.
[0037]
In addition, the present apparatus can be used in various ways by setting the reaction process, which is a processing process, and various displays such as the progress of the reaction process on the screen, and operating the keyboard, mouse or switch, or mounting a floppy or CD. Input means 6 for designating combinatorial synthesis, storage means 7 for storing various data such as dispensing amounts of various reagents, dispensing positions, dispensing timing, and the like, and communication means 8 for transmitting data Have These devices and means are connected to each other through a bus or the like.
The input means 6 and the storage means 7 correspond to the designation means.
[0038]
FIG. 2 is a top view of the processing stage 2. In the figure, reference numeral 10 denotes a group of six nozzles provided in the pipette means 1 and is equipped with a detachable dispensing tip. The nozzle is provided with a liquid level sensor (not shown) for irradiating light to detect the liquid level and receiving the reflected light, and a pressure sensor for detecting the pressure in the nozzle.
When the chemical reaction used causes an adverse effect on the environment, or when processing is performed in various atmospheres such as a gas other than air or vacuum, the pipette means and the processing stage are attached to the sealed container 9. Store and seal.
[0039]
In the present processing stage 2, the same dispensing reagent containers 11 and 13 in which the portions into which the dispensing tips attached to the nozzle group 10 used when the nozzle group 10 dispenses the same reagent enter are not partitioned from each other. Is provided. In the case of the same dispensing reagent container of reference numeral 13, a larger volume of reagent can be accommodated than in the case of reference numeral 11. Which one is used is selected based on the content of the treatment process such as the amount of the target compound group.
[0040]
The processing stage 2 is used when the six nozzle groups 10 dispense different reagents, and the different dispensing methods in which the portions into which the dispensing tips attached to the nozzles of the nozzle group 10 enter are separated from each other. Reagent containers 12 and 14 are provided. In the case of the different dispensing reagent container of reference numeral 14, it is selected when the capacity is larger than that of the container of reference numeral 12.
[0041]
The processing stage 2 contains a tip holder 15 that holds a dispensing tip attached to and removed from the nozzle group 10 and a cleaning liquid for cleaning the dispensing nozzle that has performed reagent dispensing. The reagent dispensing nozzle cleaning tank 16, the reaction nozzle cleaning tank 17 containing a cleaning liquid for cleaning the nozzle of the reacted compound, and a group of containers are arranged in a matrix of 6 rows × 6 columns to perform the reaction. Reaction plate 18 and a mixing plate 19 for mixing the reagent and magnetic particles.
[0042]
The reaction plate 18 and the mixing plate 19 are independently provided on a rotary stage 20 that can be rotated in both forward and reverse directions at an angle of 90 °, that is, can be transposed to exchange rows and columns. The rotary stage 20 corresponds to one of the moving means 3.
[0043]
In the same dispensing reagent containers 11 and 13 and the different dispensing reagent containers 12 and 14, the opening of the container is covered with a lid except for the part into which the dispensing tip enters. This prevents reagent evaporation and contamination. On the other hand, the reagent dispensing nozzle cleaning tank 16, the reaction nozzle cleaning tank 17, the reaction plate 18, and the mixing plate 19 are not provided with a lid.
[0044]
Preferably, by covering the container group portion of the processing stage 2 with a laminate sealing film, the container group is hermetically sealed, for example, an atmosphere filled with nitrogen or the like is maintained, and reagent evaporation and contamination can be reliably prevented. it can. When a laminate sealing film is used, a piercing method is used in which holes are made in the film. The dispensing tip is formed of, for example, polypropylene (PP), stainless steel (SUS), or Teflon, and is suitably formed in a needle shape when the piercing method is adopted.
[0045]
FIG. 3 is an exploded perspective view showing the same dispensing reagent container 11 detachably provided on the processing stage 2 shown in FIG. 2 in an enlarged manner. The same dispensing reagent container 11 includes a box-shaped container portion 11a formed of PP, and a substantially rectangular lid formed of PP that is detachably fitted to and covers the opening 11b of the container portion 11a. It has a body 11c.
[0046]
The container part 11a is formed of a side part 11d and an inner bottom part 11g. Here, the inner bottom portion 11g has an inclined surface 11e that falls toward the center of the inner bottom portion 11g, and is narrower than the diameter of the tip portion of the dispensing tip (not shown) and extends along the center of the inner bottom portion 11g. And a groove 11f provided.
[0047]
As a result, even when the tip of the dispensing tip comes into contact with the inner bottom surface of the container 11, almost all of the reagent in the container can be sucked, and the reagent can be diffused when the sucked reagent is discharged into the container. Can be averaged to greatly improve the stirring efficiency. The side surface 11e of the side surface portion 11d in the longitudinal direction is formed in a rectangular shape so that the lower end thereof is horizontal in order to stably support the container 11 so as not to fall down, and the container is held at the upper end. A handle 11h is provided.
[0048]
A substantially rectangular recess is provided in the center of the lid 11c, and six cross-shaped or pore-shaped cuts 11i are provided at predetermined intervals in the recess. This corresponds to the pipette means 1 provided with six, three, two, or single dispensing tips, and the dispensing tips can penetrate the cuts 11i and enter the inside of the container. Providing this cut 11i wipes the liquid in the dispensing tip, prevents evaporation of the liquid in the container, and helps prevent contamination. When storing the reagent, a rubber plate 11j is detachably fitted into the recess at the center of the lid 11c in order to completely prevent evaporation of the liquid.
The cuts and pores may be provided in the rubber plate 11j, or may be penetrated by a dispensing tip without being provided.
[0049]
FIG. 4 shows the same dispensing reagent container 21 having a smaller capacity than the same dispensing reagent container 11 shown in FIG. The container 21 includes an elongated box-shaped container portion 21a formed of PP, and an elongated rectangular lid body 21c formed of PP that is detachably fitted to and covers the opening 21b of the container portion 21a. Have. The container portion 21a is formed of a side surface portion 21d and an inner bottom portion 21g. Here, the inner bottom portion 21g is a wedge bottom, and is wider than the diameter of the inclined surface 21e inclined so as to drop toward the center of the inner bottom portion 21g and the tip of the dispensing tip (not shown). It is narrow and has a groove portion 21f provided along the center of the inner bottom portion 21g.
[0050]
The bottom surface of the groove 21f has a corrugated curved surface in which a plurality of (in this example, twelve) arcuate recesses are connected. As a result, even when the tip of the dispensing tip comes into contact with the inner bottom surface of the container 21, almost all of the reagent in the container can be sucked, and the reagent can be diffused when the sucked reagent is discharged into the container. Can be averaged to greatly improve the stirring efficiency. The side surface of the side surface portion 21d in the longitudinal direction is formed in a rectangular shape so that the lower end thereof is horizontal in order to stably support the container 21 so that the container 21 does not fall down. The handle 21h is provided.
[0051]
A substantially rectangular recess is provided in the center of the lid 21c, and twelve cross-shaped or pore-shaped cuts 21i are provided in the recess at positions corresponding to the respective arc-shaped recesses of the groove 21f. ing. This corresponds to the pipette means 1 provided with a 12-, 6-, 4-, 3-, 2- or single-dispensing tip, and the dispensing tip enters the inside of the container through the notch 21i. be able to.
[0052]
By providing this notch 21i, the liquid adhering to the outside is wiped off when the dispensing tip is inserted, and cross contamination is prevented, and evaporation of the liquid in the container is prevented. Help prevent.
When storing the reagent, a rubber plate 21j is detachably fitted into the recess in the center of the lid 21c in order to completely prevent the liquid from evaporating.
It should be noted that the rubber plate 21j may be provided with cuts or pores, or may be penetrated by a dispensing tip without being provided.
[0053]
FIG. 5 shows a modification of the same dispensing reagent container 21 shown in FIG.
The same dispensing reagent container 31 has an elongated box-shaped container portion 31a formed of PP and an elongated rectangular shape formed of PP that is detachably fitted to and covers the opening 31b of the container portion 31a. And a lid 31c. The container 31a and the lid 31c are flexibly coupled by a thinly formed hinge 31d, and when the hinge 31d is bent, the lid 31c is detachably formed so as to fit into the opening 31b. Has been.
[0054]
The container portion 31a is formed of a side surface portion 31e and an inner bottom portion 31h. Here, the inner bottom portion 31h has a wedge bottom and is inclined along the center of the inner bottom portion 31h so as to be inclined downward, and along the center of the inner bottom portion 31h. It has a groove part 31g provided narrower than the diameter of the tip part of the tip (not shown).
[0055]
The bottom surface of the groove 31g is formed with a zigzag corrugated surface having a plurality of (in this example, 12) recesses. As a result, even when the tip of the dispensing tip comes into contact with the inner bottom surface of the container 31, almost all of the reagent in the container can be sucked and the reagent can be diffused when the sucked reagent is discharged into the container. Can be averaged to greatly improve the stirring efficiency.
[0056]
On the periphery of the opening 31b at the upper end of the container part 31a, a plate larger than the side part 31e of the container part 31a and capable of being gripped at both ends in the longitudinal direction is projected. A plate 31i is provided. The upper plate 31i is supported by a support portion 31j protruding from the side surface of the side surface portion 31e on the lower surface of the portion protruding in the longitudinal direction.
[0057]
An elongated rectangular hole is formed in the center of the lid 31c, and a rubber plate 31k is fitted in the hole. Twelve cross cuts 31l are provided at positions corresponding to twelve corrugated recesses on the bottom surface of the groove 31g so that a dispensing tip is inserted through the rubber plate 31j fitted in the lid 31c. It has been. This corresponds to the pipette means 1 provided with twelve, six, four, three, two, or single dispensing tips.
It should be noted that the rubber plate provided on the container lid described above may not be provided with a cut, but may be perforated with a tip of the pipette means, or may not be able to penetrate. In this case, since there is no notch, the inside of the container is completely sealed.
[0058]
According to the present embodiment, since the lid is provided, a situation where the lid and the container are separated is prevented, and therefore, cross contamination is prevented.
[0059]
FIG. 6A shows a cross section of the dispensing tip holding part 15 provided in the processing stage 2, and FIG. 6B shows a cross sectional view of the cleaning tanks 16 and 17. Each of the washing tanks 16 and 17 has a washing dish 16a in which a dispensing pipette is aspirated and discharged, and a receiving dish 16b for collecting the washing liquid overflowing from the washing dish 16a.
[0060]
Further, as shown in the figure, a supply path 16c and a pump 16d for supplying the cleaning liquid to the cleaning dish 16a, a storage tank 16e for storing the cleaning liquid, and a waste path for discarding the cleaning liquid provided in the receiving tray 16b. 16f.
In addition, the processing stage 2 may be provided with a thermostatic device for storing the reagent or promoting the reaction, and a liquid level sensor for detecting the liquid level.
[0061]
Next, various cases in which combinatorial synthesis and structural analysis are performed using the chemical reaction processing apparatus according to the first embodiment will be described with reference to FIGS.
First, as a second embodiment, a tripeptide is finally generated by using three kinds of amino acids (A, B, C) by split synthesis based on FIG. 7 and FIG. An example of synthesis as a compound will be described. In this case, it is assumed that the magnetic force means is disposed at least outside the liquid passage communicating with each nozzle of the pipette means 1.
[0062]
As shown in FIG. 7, in step S1, processing process information is input from the input means 6 in order to specify the content of the combinatorial synthesis processing process. Specifying the treatment process, the point of split synthesis, the position of the container containing the reagent, the position of the container containing the magnetic particles, the position of the container where the reaction is performed, the control of the movement of the pipette means or the processing stage, The reaction time, the order of dispensing, etc. are also automatically specified or set based on the information stored in the storage means 7 in advance. As a result, the necessary reagent arrangement and the like are determined by the control means 4.
[0063]
In step S2, since the process is split composition, the process automatically proceeds to step S3. In step S3, the nozzle group 10 of the pipette means 1 moves to the tip holder 15 of the processing stage 2 and automatically attaches three of the six dispensing tips. The nozzle group 10 is moved to the reagent container 12, and the reagent containing the target substances amino acids A, B, and C contained in the container 12 is sucked and used for the reaction as shown in FIG. Plate 18₁Among the container groups arranged in a matrix of 6 rows × 6 columns, the amino acids are discharged and arranged in a container group of 3 rows × 1 column.
[0064]
In step S4, the nozzle group 10 to which the dispensing tip is mounted moves to the same dispensing reagent container 11 and sucks the suspension of magnetic particles from the container 11 in which the magnetic particles are stored. . Here, a substance that specifically reacts with the amino acids A, B, and C is coated or held on the surface of the magnetic particles. At that time, depending on the amino acids A, B, and C, a plurality of types of magnetic particles having different coated substances may be used.
[0065]
Next, the amino acids A, B, and C are moved to the wells of 3 rows × 1 column of the plates containing the amino acids A, B, and C, respectively, and the magnetic particles are placed in the containers containing the reagents containing the amino acids A, B, and C It dispenses and mixes, and each amino acid A, B, C reacts with the magnetic particles and binds. At this stage, each of the 3 rows × 1 column containers accommodates the combined body of magnetic particles-A, magnetic particles-B, and magnetic particles-C.
[0066]
In step S5, the reaction plate 18 of FIG.₁The mixed liquid containing the conjugate of amino acid A, B, C and magnetic particles contained in the three containers is sucked by the nozzle group 10 to which a dispensing tip is attached, and is moved by the moving means 3. Move to the mixing plate 19 of the processing stage 2 and discharge each mixture at the same time to mix and randomize the suspension of magnetic particles and conjugates of each amino acid A, B, C, A mixed solution in which equal amounts of the combined bodies are uniformly mixed is generated.
[0067]
In step S6, the mixed solution in which the combined body of amino acids A, B, C and magnetic particles mixed in the mixing plate 19 is uniformly suspended is again equalized by the nozzle group 10 in triplicate. As shown in FIG. 8, the reaction plates 18 are sucked one by one, and each of the sucked mixed solutions is moved by the moving means 3.₁1/3 of the liquid mixture sucked for each nozzle (first amino acids A, B, and 1) while being translated into nine container groups of 3 columns × 3 rows, which is a partial matrix of a matrix of 6 columns × 6 rows of (When the amount of C is 1) is distributed equally to a total of 9 container groups. At that time, since the number of rows and columns is the same, the plate does not need to be rotated 90 ° and transposed.
[0068]
In step S7, the nozzle group 10 again aspirates each of the reagents each containing the amino acids A, B, and C contained in the different dispensing reagent container 12 one by one. Each nozzle is translated into a container group of 1 column x 3 rows by dividing it into 1/3 each element of the matrix matrix of rows x 3 columns, thereby dividing and dispensing equal amounts of different amino acids for each column.
[0069]
Therefore, each container of the matrix of 3 rows × 3 columns in FIG. 8 is mixed with a reagent containing amino acids A, B and C, a mixture of magnetic particles and a conjugate of amino acids A, B and C. It will be.
[0070]
In step S8, a conjugate in which two kinds of amino acids are bound is generated by reaction and incubation between amino acids.
Therefore, in the first row of container groups, magnetic particles-AA, magnetic particles-BA, magnetic particles-CA, and in the second row of containers, magnetic particles-AB, Magnetic particle-BB, magnetic particle-CB, and in the third row of containers, each combination of magnetic particle-A-C, magnetic particle-BC, magnetic particle-CC A group of compounds consisting of bodies is obtained.
[0071]
In step S9, the nozzle group aspirates the entire amount from each of the matrix-like container group of 3 columns × 3 rows for each row, and discharges them to one container of the mixing plate 19, and the compound A uniform mixture of groups is produced. In the mixed solution, the respective conjugates are mixed uniformly.
[0072]
In step S10, the nozzle group 10 sucks the mixed solution from the mixing plate 19 in nine equal portions, and the reaction plate 18 shown in FIG.₁, 18₂Of the mixed solution containing the conjugate of amino acids A, B, and C for each row by moving the nozzle group 10 in parallel to each container group in a matrix of 3 columns × 6 rows and 3 columns × 3 rows. Dispense in 1/27 of the total volume, that is, in 2/9 equal portions.
[0073]
In step S11, the nozzle group 10 further sucks amino acids A, B, and C from the different dispensing reagent container 12 one by one for each nozzle, and translates the nozzle group 10 for each row. While dividing into 1/9 equal parts, the mixture is mixed with the liquid mixture contained in each well. Also in this case, the rotary stage 20₁, 20₂There is no need to rotate 90 ° and transpose it.
[0074]
In step S12, a conjugate in which three kinds of amino acids are combined is generated by reaction and incubation between amino acids. Accordingly, the following combinations are included in each well of 3 columns × 6 rows for each column. For the first row, magnetic particles-AAA, magnetic particles-BAA, magnetic particles-CAA, magnetic particles-ABAA, magnetic particles- B-B-A, magnetic particles-C-B-A, magnetic particles-A-C-A, magnetic particles-B-C-A, and magnetic particles-C-C-A.
[0075]
For the second row, magnetic particles -A-A-B, magnetic particles-B-A-B, magnetic particles-C-A-B, magnetic particles-A-B-B, magnetic particles- B-B-B, magnetic particles-CBB, magnetic particles-A-C-B, magnetic particles-B-C-B, and magnetic particles-C-C-B.
[0076]
For the third column, magnetic particles -A-A-C, magnetic particles-B-A-C, magnetic particles-C-A-C, magnetic particles-A-B-C, magnetic particles- B-B-C, magnetic particles-C-B-C, magnetic particles-A-C-C, magnetic particles-B-C-C, and magnetic particles-C-C-C. At this stage, a conjugate in which all 27 types of tripeptides are bound to magnetic particles is obtained.
[0077]
In step S13, a dissociating agent is dispensed into the well group which is a container group arranged in a matrix of 3 columns × 6 rows and 3 columns × 3 rows to dissociate the conjugates from the magnetic particles. Each tripeptide can be obtained by separating only the magnetic particles.
[0078]
Subsequently, as a third embodiment, based on FIG. 7 and FIG._i-B_j-C_kAn apparatus in which at least magnetic means (not shown) is disposed outside each container (for reaction) of the processing stage 2 and can adsorb and desorb magnetic particles on the inner surface of each container. A case where the parallel synthesis method is applied when used will be described.
[0079]
In step S1 of FIG. 7, the operator inputs processing process information from the input means 6 of FIG. 1 in order to specify the processing process of combinatorial synthesis. When the process is specified, the point of parallel synthesis, the position of the reagent storage container, the position of the magnetic particle storage container, the position of the container where the reaction is performed, the control of the movement of the pipette means or the processing stage, the reaction time, the dispensing The order and the like are also automatically specified or set based on information stored in the storage unit 7 in advance.
[0080]
In step S2, since the process is a parallel synthesis method, the process automatically proceeds to step S21.
In step S21, the nozzle group 10 of the pipette means 1 moves to the tip holder 15 of the processing stage 2 and automatically attaches three of the six dispensing tips. The target substance A contained in the container 12 is moved by moving the nozzle group 10 to the dispensing reagent container 12.₁, A₂, A_ThreeAs shown in FIG. 9, the reaction plate 18 is separately sucked by each nozzle of the nozzle group 10 with each reagent separately contained.₁Among the container groups arranged in a matrix of 6 rows × 6 columns, the reagent is discharged into a container group of 3 rows × 1 column.
[0081]
In step S22, the nozzle group 10 to which the dispensing tip is attached moves to the same dispensing reagent container 11 and sucks the suspension containing the magnetic particles from the container 11 in which the magnetic particles are stored. To do. Here, on the surface of the magnetic particles, the target substance A₁, A₂, A_ThreeA substance that specifically reacts with is coated. At that time, target substance A₁, A₂, A_ThreeDepending on the case, a plurality of types of magnetic particles having different coated substances may be used.
[0082]
Next, the target substance A₁, A₂, A_ThreeAre moved to the wells of 3 rows × 1 column of the plates in which the magnetic substance particles are stored.₁, A₂, A_ThreeIs dispensed into each container containing each reagent, and each target substance A₁, A₂, A_ThreeAre combined with magnetic particles to be combined with magnetic particles. At this stage, the container group of 3 rows × 1 column is sequentially filled with magnetic particles-A.₁, Magnetic particles-A₂, Magnetic particles-A_ThreeThus, a liquid containing each of the above conjugates is accommodated.
[0083]
In step S23, the nozzle group 10 draws in each reagent including the conjugate bound to the magnetic particles, and discharges them in equal amounts while moving in translation by 1/3 of the original amount. Plate 18₁A different liquid is dispensed for each column into a group of containers arranged in a matrix of 6 rows × 6 columns. Thereafter, a magnetic field is applied to the inside of the container by magnetic means individually provided on the outside of each container, and the combined body combined with the magnetic particles is attracted and held on the inner surface of each container. Thereafter, the pipette means 1 sucks out the stored liquid excluding the bonded body held by adsorption on the inner surface, removes it by transferring it to another container for discharge, and then separating the magnetic means from the container. Or by turning off the electromagnet.
[0084]
In step S24, the moving means 3 moves the reaction plate 18 as shown in FIG.₁The rotary stage 20 on which is mounted₁Is rotated 90 ° to transpose the rows and columns of the container group arranged in a matrix.
[0085]
In step S25, after washing the dispensing tip in the washing tank 16, or after replacing the dispensing tip with the tip holder, the target substance B is removed from the container 12 for different dispensing reagents.₁, B₂, B_ThreeIn contrast to the transposed matrix container group in which the row and column are exchanged by aspirating 1 each of the reagents separately containing 1 and aspirating 1/3 or 1/3, respectively, the nozzle group 10 A that is bonded to the magnetic particles already contained in each well by translation₁~ A_ThreeAre sequentially dispensed into a container in which each of the conjugates is housed.
[0086]
In step S26, by the reaction and incubation, at this stage, the three containers in the first row of the transposition matrix are each filled with magnetic particles-A.₁-B₁, Magnetic particles-A₁-B₂, Magnetic particles-A₁-B_ThreeIn the three containers in the second row of the transposition matrix, the magnetic particles -A are respectively stored.₂-B₁, Magnetic particles-A₂-B₂, Magnetic particles-A₂-B_ThreeThe liquid containing the combined body is stored, and the three containers in the third row each have magnetic particles-A._Three-B₁, Magnetic particles-A_Three-B₂, Magnetic particles-A_Three-B_ThreeEach of the liquids containing all nine types of conjugates is accommodated.
[0087]
In step S27, the nozzle group 10 moves in a translational manner to sequentially suck all contained liquids containing the conjugates three by three from the container group of the transposed matrix from the container. Different combinations for each column in the 9 types of combined bodies in a total of 6 columns × 3 rows and 3 columns × 3 rows of containers of matrix containers transposed by 3 each, ie 1/9 of the initial quantity The liquid containing the body is dispensed, and the three containers belonging to each row are dispensed while being translated so that the liquid containing the same conjugate is dispensed. After that, the magnetic body provided on the outside of each container acts on the inside of the container to adsorb and hold the bonded body combined with the magnetic particles on the inner surface of each container, and the pipette means 1 adsorbs on the inner surface. The accommodated liquid excluding the held combined body is sucked and removed by being transferred to another discharge container and discharged, and then the action of the magnetic field by the magnetic force means is removed.
[0088]
In step S 28, the reaction plate 18 is moved by the moving means 3 as shown in FIG.₁And 18₂Each rotary stage 20₁, 20₂Is reversed 90 ° to return the transposed matrix row and column to the original matrix row and column.
[0089]
In step S29, after the dispensing tip is washed in the washing tank 16, or after the dispensing tip is replaced by the tip holder, the target substance C is removed from the different dispensing reagent container 12.₁, C₂, C_ThreeAspirate each 1 quantity from each reagent separately and aspirate 1/9 quantity or 1/9 quantity to collect 3 x 6 matrix matrix and 3 x 3 matrix By moving the nozzle group 10 in translation with respect to the container group, the conjugate with the magnetic particles already contained in each well is sequentially dispensed.
[0090]
In step S30, the nine particles (6 and 3) in the first row of the matrix are each filled with magnetic particles-A by reaction and incubation.₁-B₁-C₁, Magnetic particles-A₁-B₂-C₁, Magnetic particles-A₁-B_Three-C₁, Magnetic particles-A₂-B₁-C₁, Magnetic particles-A₂-B₂-C₁, Magnetic particles-A₂-B_Three-C₁, Magnetic particles-A_Three-B₁-C₁, Magnetic particles-A_Three-B₂-C₁, Magnetic particles-A_Three-B_Three-C₁9 types of conjugates are obtained, and one type each is accommodated.
[0091]
The nine containers (6 and 3) in the second row of the matrix have magnetic particles-A, respectively.₁-B₁-C₂, Magnetic particles-A₁-B₂-C₂, Magnetic particles-A₁-B_Three-C₂, Magnetic particles-A₂-B₁-C₂, Magnetic particles-A₂-B₂-C₂, Magnetic particles-A₂-B_Three-C₂, Magnetic particles-A_Three-B₁-C₂, Magnetic particles-A_Three-B₂-C₂, Magnetic particles-A_Three-B_Three-C₂Each of these nine types of conjugates is accommodated.
[0092]
The nine containers (6 and 3) in the second row of the matrix have magnetic particles-A, respectively.₁-B₁-C_Three, Magnetic particles-A₁-B₂-C_Three, Magnetic particles-A₁-B_Three-C_Three, Magnetic particles-A₂-B₁-C_Three, Magnetic particles-A₂-B₂-C_Three, Magnetic particles-A₂-B_Three-C_Three, Magnetic particles-A_Three-B₁-C_Three, Magnetic particles-A_Three-B₂-C_Three, Magnetic particles-A_Three-B_Three-C_ThreeEach of these nine types of conjugates is accommodated. In this way, a total of 27 types of conjugate groups are obtained.
[0093]
In step S31, a dissociating agent is dispensed into the wells arranged in a matrix of 3 columns × 6 rows and 3 columns × 3 rows to dissociate the conjugates from the magnetic particles. Can be obtained to obtain the final product compound.
[0094]
Subsequently, as a fourth embodiment, a labeling process for performing a structural analysis using a chemical reaction processing apparatus using a pipette means provided with the magnetic force means will be described.
As shown in FIG. 7, in step S <b> 1, processing process information is input from the input unit 6 in order to specify the content of the structural analysis processing process. When the process is specified, structural analysis is performed, the position of the container in which the reagent is stored, the position of the container in which the magnetic particles are stored, the position of the container in which the reaction is performed, the movement of the pipetting means or the processing stage The reaction time, the order of dispensing, etc. are also automatically specified or set based on information stored in the storage means 7 in advance.
[0095]
In step S2, since the process is a structural analysis, the process automatically proceeds to step S51. In step S51, the nozzle group 10 of the pipette means moves to the tip holder 15 of the processing stage 2 and automatically attaches three consecutive dispensing tips to the different dispensing reagent container 12 of the processing stage 2. The nozzle group 10 is moved to suck the reagent containing amino acids A, B, and C accommodated in the container 12, and as shown in FIG. 8, the reaction plate 18 has a matrix of 6 rows × 6 columns. Among the arranged container groups, the amino acids A, B, and C are dispensed into a container group of 3 rows × 1 column.
[0096]
In step S52, in order to identify the amino acid, amino acid A is changed to T.₁, Amino acid B has T₂, Amino acid C has T₁+ T₂In order to correspond to the tags, the tags stored in another different dispensing reagent container 12 are aspirated and dispensed into the container.
[0097]
In step S53, the magnetic pair particles coated with the amino group that binds to the amino acid and the tag by reaction are dispensed and mixed into a container in which the amino acid and the tag are dispensed. At that time, the dispensing amount of the tag is set so that the tag is bound to 0.5 to 1% of the amino group and the amino acid is bound to the remaining amino group.
[0098]
Therefore, at this stage, each of the three containers contains magnetic particles-amino acid A-tag T.₁, Magnetic particles-amino acid B-tag T₂, Magnetic particles-amino acid C-tag T₁+ T₂Are obtained.
[0099]
In step S54, the magnetic particles are mixed and randomized with each conjugate having amino acids and tags bound thereto.
In step S55, an equal amount is divided into three containers and the amino protecting group is cleaved.
[0100]
In step S56, a new tag T is added to the released amino acid._Three, T_Four, T_Three+ T_FourIs dispensed into each of three containers. Where tag T_ThreeIdentifies amino acid X and tag T_FourIdentifies amino acid Y and tag T_Three+ T_FourIs for identifying the amino acid Z.
[0101]
In step S57, amino acids X, Y, and Z are dispensed into the three containers, respectively, by the pipette means.
In step S58, amino acids are combined by reaction and incubation. At this stage, the first container has T₁T_Three-Magnetic particles-AX, T₂T_Three-Magnetic particles-BX, T₁T₂T_Three-Magnetic particles-C-X, T in the second container₁T_Four-Magnetic particles-AY, T₂T_Four-Magnetic particles-BY, T₁T₂T_Four-Magnetic particles-C-Y, T in the third container₁T_ThreeT_Four-Magnetic particles-AZ, T₂T_ThreeT_Four-Magnetic particles-BZ, T₁T₂T_ThreeT_Four-Magnetic particles-CZ are accommodated, and the conjugate of each amino acid is labeled.
[0102]
Subsequently, after the assay, the tag is cleaved from the extracted magnetic particles in step S59, and then analyzed by EC-GC in step S60, whereby the amino acid sequence bound to the magnetic particles is analyzed. Can be specified.
[0103]
Subsequently, as a fifth embodiment, based on FIGS. 10 and 11, at least magnetic force means is arranged outside the liquid passage communicating with each nozzle of the pipette means 1 by parallel synthesis method (paralel synthesis). Another example will be described in which a tripeptide is synthesized as a final product compound using a reagent containing three kinds of amino acids (A, B, and C).
[0104]
In step S1 in FIG. 7, processing process information is input from the input means 6 in order to specify the content of the combinatorial synthesis processing process. When the processing process is specified, in step S2 in FIG. 7, parallel synthesis is performed, the position of the container in which the reagent is stored, the position of the container in which the magnetic particles are stored, the position of the container in which the reaction is performed, and pipette means Alternatively, the control of the movement of the processing stage, the reaction time, the order of dispensing, etc. are automatically specified or set based on the information stored in the storage means 7 in advance. As a result, the necessary reagent arrangement and the like are determined by the control means 4.
[0105]
According to this analysis, in this embodiment, the final product with the number of component bonds of 3 is generated using the three types of substance groups (A, B, C). Using symbols, n = 3, m = 9, k = 3. That is, 3 groups of 3 columns × 9 rows are required. However, since only 6 columns × 6 rows of matrix container groups are arranged on one plate, each 3 columns × 6 rows container group is arranged across two plates. In this embodiment, the substance group is arranged as follows.
[0106]
First, the nozzle group 10 of the pipette means 1 moves to the tip holder 15 of the processing stage 2 and automatically attaches three of the six dispensing tips to the different dispensing reagent container of the processing stage. The nozzle group 10 is moved to 12, the liquid containing the amino acids A, B, and C contained in the container 12 is sucked, and the reaction plate 18 in FIG.₁, 18₂And the reaction plate 18 in FIG._Three, 18_FourThe amino acids A, B, and C are respectively dispensed and arranged in a matrix in four groups of containers arranged in a matrix of 6 rows × 6 columns. Here, it is assumed that the nozzle group 10 is arranged in a vector of 1 column × 3 rows.
[0107]
In addition, each reaction plate 18₁, 18₂, 18_Three, 18_FourAre each rotary stage 20₁, 20₂, 20_Three, 20_FourIs provided. Reaction plate 18₁In the above, each container group of 6 columns × 6 rows is dispensed in the order of A, B, and C, and each matrix of 3 columns × 6 rows is stored in a container group M.₁, Container group M₂It expresses. Reaction plate 18₂In the above, each container group in each column of 6 columns × 3 rows is arranged in the order of A, B, C, and each matrix of 3 columns × 3 rows is stored in each container group M._Three, Container group M_FourIt expresses. Further, the reaction plate 18 in FIG._ThreeAbove, it arranges in order of A, B, C for each container group of each column of 3 columns × 6 rows, and the container group M_FiveIt expresses. Reaction plate 18 in FIG._FourAbove, container groups M are arranged in the order of A, B, C for each container group of 3 columns × 3 rows.₆It expresses. That is, container group M₁And M_ThreeCorresponds to the first group of 3 columns × 9 rows of containers, and the container group M₂And M_FourIs equivalent to the 3 column × 9 row container group of the second group, and the container group M_FiveAnd M₆Is equivalent to the 3 column × 9 row container group of the third group.
[0108]
Next, the reaction plate 18 corresponding to the first group of 3 columns × 9 rows of containers.₁3 column x 6 row container group M₁And reaction plate 18₂3 column x 3 row container group M_ThreeOn the other hand, by the nozzle of the pipette means 1, an equal amount of a liquid in which magnetic particles coated or held with a substance that specifically reacts and binds to A, B, and C are suspended is dispensed. Then, the magnetic particles and the amino acids A, B, and C are bonded to form a bonded body.
[0109]
Next, a container group M corresponding to a 3 column × 9 row container group of the first group in which a conjugate of magnetic particles and each amino acid is generated and accommodated.₁And container group M_ThreeThe reaction plate 18 corresponding to the second group of 3 columns × 9 rows of containers, with the transport source as the transfer source and only the combined body as the transfer target.₁, 18₂Container group M₂And M_FourAs the transfer destination, the rotary stage 20₁, 20₂Is transferred without transposition.
[0110]
Each of the three nozzles of the pipette means 1 is first placed in the container group M.₁In the first column of (M₁)₁₁, (M₁)₁₂, (M₁)₁₃Are translated into three containers (subscripts represent column and row numbers). When sucking and discharging all the contained liquid from the container group that is the transfer source, the combined body is separated by applying a magnetic field to the liquid passage by the magnetic force means, and the container group M at the transfer destination is separated.₂In the first column of (M₂)₁₁, (M₂)₁₂, (M₂)₁₃The pipette means 1 is translated into the three containers. Subsequently, the three containers (M₂)₁₁, (M₂)₁₂, (M₂)₁₃Then, the combined liquid is transferred from the pipette means 1 to each container by sucking and discharging the contained liquid without applying a magnetic field by the magnetic means to the liquid passage.
[0111]
Similarly, the nozzle group 10 of the pipette means 1 is connected to the container group M in which the combined body is accommodated.₁In the first column (M₁)₁₄, (M₁)₁₅, (M₁)₁₆Translated into three containers. When sucking and discharging all the contained liquid from the container group that is the transfer source, the combined body is separated by applying a magnetic field to the liquid passage by the magnetic force means, and the container group M at the transfer destination is separated.₂In the second column of (M₂)_{twenty one}, (M₂)_{twenty two}, (M₂)_{twenty three}The pipette means 1 is translated into the three containers. Subsequently, the three containers (M₂)_{twenty one}, (M₂)_{twenty two}, (M₂)_{twenty three}Then, the combined liquid is transferred from the pipette means 1 to each container by sucking and discharging the contained liquid without applying a magnetic field by the magnetic means to the liquid passage.
Next, the nozzle group 10 of the pipette means 1 is moved to the container group M in which the combined body is accommodated._ThreeIn the first column (M₁)₁₄, (M₁)₁₅, (M₁)₁₆Translated into three containers. In the same manner, the combined body is separated from the container group that is the transfer source, and the container group M that is the transfer destination.₂In the third column (M₂)₃₁, (M₂)₃₂, (M₂)₃₃The nozzle group 10 of the pipette means 1 is translated into the three containers, and the combined body is transferred from the pipette means 1 to each container in the same manner.
[0112]
In the same manner, M is set so that the three container groups are placed together as a transposition position on a virtual matrix that is made into one element.₁And M_ThreeFrom the first group of 3 columns × 9 rows container group, M₂And M_FourWhen only the conjugate is transferred to the second group of 3 columns × 9 rows container group, the conjugate and the reagent are transferred to the container group M as shown in FIG.₂And M_FourThe second group is arranged in a 3 column × 9 row container group.
That is, at this stage, by the reaction and incubation, magnetic particles-AA, magnetic particles-AB, magnetic particles-AC, magnetic particles-BA, magnetic particles-BB, Magnetic particles-BC, magnetic particles-CA, magnetic particles-CB, and magnetic particles-CC are produced.
[0113]
Subsequently, the reaction plate 18₁And 18₂Rotating stage 20 provided with₁And 20₂Is rotated 90 ° and transposed, as shown in FIG. 11C, the reaction plate 18₁And reaction plate 18₂2nd 3 columns × 9 rows container group M₂And M_FourAre transposed and each container group M₂ ^TAnd M_Four ^T("T" represents a transposed matrix).
[0114]
Next, the nozzle group 10 of the pipette means 1 is, for example, a container group M₂3 containers (M₂ ^T)₆₁, (M₂ ^T)₆₂, (M₂ ^T)₆₃When the entire amount is sucked, a magnetic field is applied to the liquid passage of the pipette means 1 by a magnetic means provided outside, and the combined body is separated by adsorbing and holding on the inner wall of the liquid passage of the pipette means. When the combined body is separated, the reaction plate 18 is a 3 column × 9 row container group of the third group of the transfer destination._ThreeContainer group M_FiveIn the first column of (M_Five)₁₁, (M_Five)₁₂, (M_Five)₁₃Move to three containers. Subsequently, the (M_Five)₁₁, (M_Five)₁₂, (M_Five)₁₃From the three containers, the combined liquid is transferred from the pipette means 1 to each container by sucking and discharging the contained liquid without applying a magnetic field by the magnetic means to the liquid passage.
[0115]
Similarly, the nozzle group 10 of the pipette means 1 is connected to the container group M in which the combined body is accommodated.₂ ^TIn the fifth column (M₂ ^T)₅₁, (M₂ ^T)₅₂, (M₂ ^T)₅₃Translate to. When the entire amount of the contents of the container group as the transfer source is sucked, the magnetic force means applies a magnetic field to the liquid passage of the pipette means 1 to separate the combined body. When the conjugate is separated, the reaction plate 18 as the transfer destination._ThreeContainer group M_FiveIn the second column of (M_Five)_{twenty one}, (M_Five)_{twenty two}, (M_Five)_{twenty three}Move to three containers. Subsequently, the (M_Five)_{twenty one}, (M_Five)_{twenty two}, (M_Five)_{twenty three}From the three containers, the combined liquid held by the pipette means is transferred to each container by sucking and discharging the contained liquid without applying the magnetic field by the magnetic means to the liquid passage.
[0116]
Similarly, sequentially M₂ ^TOnly the conjugate is separated from the container group, and the container group M_FiveTo each corresponding container group and finally (M_Four ^T)₁₁, (M_Four ^T)₁₂, (M_Four ^T)₁₃The reaction plate 18_FourContainer group M₆In the third column (M₆)₃₁, (M₆)₃₂, (M₆)₃₃Will be transferred to.
[0117]
Thus, as final products, magnetic particles-AAAA, magnetic particles-AABB, magnetic particles-AAAC, magnetic particles-ABAA, magnetic particles- A-B-B, magnetic particles-A-B-C, magnetic particles-A-C-A, magnetic particles-A-C-B, magnetic particles-A-C-C, magnetic particles- B-A-A, magnetic particles-B-A-B, magnetic particles-B-A-C, magnetic particles-B-B-A, magnetic particles-B-B-B, magnetic particles- B-B-C, magnetic particles-B-C-A, magnetic particles-B-C-B, magnetic particles-B-C-C, magnetic particles-C-A-A, magnetic particles- C-A-B, magnetic particles-C-A-C, magnetic particles-C-B-A, magnetic particles-C-B-B, magnetic particles-C-B-C, magnetic particles- C-C-A, magnetic particles-C-C-B, magnetic particles-C- -C is, will be housed in 27 the vessels of the third column 3 × 9 rows container group group FIG. 11 (d).
[0118]
In the processing stage 2 described above, the rotation stage 20 is provided for each plate 18 and can be rotated for each plate 18. However, the present invention is not limited to this case. For example, in the processing stage 35 shown in FIG. A plurality of plates 18 (four plates in this example) may be placed on one rotation stage 36 so that the plurality of plates can rotate simultaneously.
In the embodiment described above, only the split synthesis method and the parallel synthesis method have been described, but a synthesis method in which both are combined may be used.
Furthermore, the synthesis method according to the present invention can be applied not only to biopolymers but also to organic substances and inorganic substances.
[0119]
In the above-described split synthesis method, mixing is performed twice and a (= 3) equal division of the mixed solution is performed twice. However, the present invention is not limited to this case, and a equal division is performed. By repeating n times aⁿIndividual target compound groups can be obtained.
In addition, the split synthesis method has shown an example of synthesizing a tripeptide as a library, but is not limited to this example. For example, not only other proteins, biopolymers of DNA and polysaccharides, but also other organic and inorganic substances The present invention can also be applied to sugar.
[0120]
In the above example, the measurement means has not been described. However, by providing the measurement means and further providing an apparatus that also analyzes the measurement result, it is possible to perform from measurement to analysis consistently.
[0121]
In addition to using one kind of magnetic particles, a necessary kind of magnetic particles is selected from two or more kinds of magnetic particles based on the contents of the treatment process. Also good. In this way, it is possible to apply by coating the magnetic particles with a substance that reacts specifically with the substance involved in the reaction, or by selecting and coating a substance that does not adversely affect the substance involved in the reaction. The range of possible treatments is increased and versatile and reliable chemical reaction treatment can be performed.
Furthermore, for the convenience of explanation, the examples of the substance group used in each of the above-described embodiments are merely examples described, and the contents of the chemical reaction process, the type and number of target substances, the final Products and the like are not limited to these examples.
[0122]
【The invention's effect】
As described above, in the first invention, the pipette means provided with the magnetic force means or the moving means that enables translocation movement or translational movement of a part or the whole of the processing stage is provided, and designation by the designation means. Based on the arrangement of the substance group such as the reagent, the transfer object, the transfer source and the transfer destination container or container group of each container, the transfer movement or translational movement at the time of transfer, and the pipette means Control means for specifying and instructing the operation is provided.
[0123]
Therefore, by arranging a large number of substance groups in a matrix and performing translocation and translational movements, chemical reaction treatment can be performed systematically and efficiently with a simple and lean operation, and the apparatus The configuration can be made simple and compact.
[0124]
In addition, by arranging magnetic means, it is possible to simplify or diversify the process by selecting and specifying a transfer target from the contents in each container, and to perform various processes with high reliability. It can be done with a compact and simple device.
By handling a large number and a small amount of material group, it is possible to efficiently produce a variety of products in small quantities.
[0125]
In the second invention, according to the content of the processing process and the like, by setting the number and non-arrangement of the multi-nozzle of the pipette means, the number and arrangement of the container group of the processing stage, efficient use without waste, The entire apparatus can be made compact.
[0126]
The third invention uses a dispensing tip that is detachably attached to the nozzle of a pipette means dispenser to perform a reliable chemical reaction process without cross-contamination without requiring cleaning. Can do.
According to the fourth aspect of the present invention, a magnetic means is provided outside a part or the whole of the container group so that the magnetic particles can be adsorbed and desorbed on the inner surface of each container.
As a result, it is possible to select and transfer a combined body with the magnetic particles or a body other than the combined body from the stored materials stored in the container, so that various treatments are possible.
[0127]
According to a fifth aspect of the present invention, by providing a lid member on the container, the evaporation of the liquid in the container, contamination, or generated gas is confined to prevent adverse effects on the environment, and the deposit on the outside of the pipette means is wiped off. Easy to handle.
[0128]
According to the sixth aspect of the present invention, chemical reaction processing can be performed consistently or efficiently by providing various devices necessary for processing in the processing stage.
According to the seventh aspect of the present invention, by providing a processing stage or the like in a sealed space, a chemical reaction harmful to the human body or the like can be contained to protect the environment, or various atmospheres, conditions or situations can be revealed in the sealed space. As a result, diverse processing can be performed.
[0129]
According to the eighth aspect of the invention, the chemical reaction treatment can be performed efficiently or quickly by reducing the treatment time and the like.
The ninth invention uses the pipette means, the processing stage, the moving means and the control means provided with the magnetic means so as to control based on the contents of the processing process.
Therefore, similarly to the effect exhibited by the first invention, the chemical reaction process can be performed easily, efficiently or quickly based on the contents of the process, and the configuration of the apparatus can be made simple and compact. .
[0130]
According to the tenth invention, processing such as a so-called split synthesis method can be performed efficiently and quickly with a simple structure and operation.
According to the eleventh aspect of the invention or the twelfth aspect of the invention, the combined body with the magnetic particles is separated from the contents contained in each container, and the contents other than the combined substance are removed while Therefore, it is possible to efficiently and quickly perform a reliable treatment by removing impurities and waste products other than the target substance.
[0131]
In the thirteenth invention, by associating the identifier with the magnetic particle, the magnetic particle, and therefore, the substance that binds to the magnetic particle can be labeled.
As a result, the structural analysis of the synthesized object can be easily and reliably performed.
[Brief description of the drawings]
FIG. 1 is a block diagram according to an embodiment of the present invention.
FIG. 2 is a plan view showing a processing stage according to the embodiment of the present invention.
FIG. 3 is a perspective view showing a first example of the same reagent dispensing container according to the embodiment of the present invention.
FIG. 4 is a perspective view showing a second example of the same reagent dispensing container according to the embodiment of the present invention.
FIG. 5 is a diagram showing a third example of the same reagent dispensing container according to the embodiment of the present invention.
FIG. 6 is a view showing a cleaning tank and a chip holder according to the embodiment of the present invention.
FIG. 7 is a flowchart according to the embodiment of the invention.
FIG. 8 is an explanatory diagram of the split synthesis method according to the embodiment of the present invention.
FIG. 9 is an explanatory diagram of a parallel synthesis method according to an embodiment of the present invention.
FIG. 10 is an explanatory diagram of another parallel synthesis method according to the embodiment of the present invention (1).
FIG. 11 is an explanatory diagram (2) of another parallel synthesis method according to the embodiment of the present invention;
FIG. 12 is a plan view showing another processing stage according to the embodiment of the present invention.
[Explanation of symbols]
1 ... Pipette means
2,35 ... Processing stage
3 ... Moving means
4. Control means
5. Display means
6 ... Input means
7. Storage means
10 ... Nozzle group
11, 13 ... Same dispensing reagent container
12, 14 ... Container for different dispensing reagents
16 ... Reagent dispensing nozzle washing tank
17 ... Reaction nozzle cleaning tank
18 ... Reaction plate
19 ... Mixing plate
20 ... Rotation stage

Claims (13)

Pipette means having a multi-nozzle capable of sucking and discharging liquid, a processing stage provided with a container group in which containers are arranged in a matrix, and a part or all of the pipette means or the processing stage container group Rows and columns of magnetic means capable of adsorbing and desorbing magnetic particles on the liquid passage of the pipette means or the inner surface of each container, and part or all of the nozzle group of the pipette means or the container group of the processing stage Moving means that enables transposition movement that moves by exchanging and translation movement that moves without exchanging rows and columns, designation means that designates the contents of combinatorial synthesis, and control means that controls the operation of each means At the same time, the control means arranges the substance group containing the magnetic particles on the container group based on the designation by the designation means, and stores the contents contained in each container. Specify the object to be transferred, the container or group of the transfer source and the transfer destination, the displacement or translational movement at the time of transfer, and the operations such as the suction of the pipette means and the adsorption by the magnetic means. A combinatorial synthesis apparatus using magnetic particles characterized by instructing each means. 2. The magnetic particles according to claim 1, wherein the arrangement of the nozzles of the pipette means or the arrangement of the containers of the processing stage is determined in relation to each other and based on the contents of the processing process. Combinatorial synthesizer . The combinatorial synthesis apparatus using magnetic particles according to claim 1 or 2, wherein the pipette means has a dispensing tip that is detachably attached to a nozzle of a dispenser of a liquid suction line. 2. A part or a whole of the container group of the processing stage is provided with a magnetic means on the outside so that magnetic particles can be adsorbed and desorbed on the inner surface of each container. 4. A combinatorial synthesizer using the magnetic particles according to any one of 3 above. A part or the whole of the container group provided on the processing stage is covered with a film-like or plate-like lid member, and can be penetrated by the tip of a dispensing tip attached to the nozzle of the pipette means. A combinatorial synthesizer using the magnetic particles according to claim 1. 2. The processing stage is provided with instruments necessary for a processing process such as a dispensing container, a cleaning container, a reaction container, a thermostat or / and a measuring apparatus. A combinatorial synthesizer using the magnetic particles according to any one of 1 to 5. The combinatorial synthesizer using magnetic particles according to any one of claims 1 to 6, wherein the processing stage and the pipette means are provided in an airtight container. The control means reduces the processing time during the processing process, the amount of movement, the movement time or the energy consumption by the moving means, or simplifies the operation with respect to the pipette means or the moving means. The combinatorial synthesizer using magnetic particles according to claim 1 or 7, characterized in that the control is efficiently performed. The processes arranged based on the contents of the designated processing steps by using the pipette means, the processing stage, the designation means, the movement means and the control means according to any one of claims 1 to 8. A group of substances such as reagents is arranged on the basis of the process (S1) of arranging a plurality of types of substance groups such as reagents in a part or the whole of the stage container group and the contents of the designated processing process. A step (S2) of dispensing a liquid containing magnetic particles in a part or the whole of the container group, and combining the target substance and the magnetic particles in each substance by reaction or the like (S2); Based on the contents of the specified processing process, transposition movement that moves by changing the row and column or translation movement that moves without changing the row and column for a part or the whole of the nozzle group of the pipette means or the container group of the processing stage By And a step (S3) of transferring a part or all of the contents stored in the transfer source container or container group to the transfer destination container or container group (S3). Combinatorial synthesis method used. The step (S1) is, a ₁ ~a _n reagent dispensing and disposed in the container group of one row and n columns × equal amounts the of the container group arranged in a matrix, b ₁ ~b _m reagent Are dispensed and arranged in an equal amount in m column × 1 row container group, and the step (S2) dispenses magnetic particles into the container group in which the reagent is arranged, and a _{1 to} a _n is a process of generating a conjugate by binding the target substance in the reagent of _n and the magnetic particles by reaction or the like, and the step (S3) pipettes the entire contents of each container containing the conjugate. The mixture is transported into a common mixing container by means, mixed, and the mixed solution is divided into m equal parts, and pipette means by an equal amount into a container group in each column and m rows of containers arranged in a matrix of n columns x m rows. as well as transported by, select the translation or transposition moves during feeding該移, reagents b ₁ ~b _m and n equal parts, before Using magnetic particles according to claim 9, characterized in that the step of transferring by n columns × m rows of a matrix in sequence containers each row n rows of containers equal amount by pipetting means groups Combinatorial synthesis method . The step (S1), the reagent of a ₁ ~a _n of the container group arranged in a matrix in equal amounts, equal amounts by dispensing and disposed in the container group n columns × m rows, b ₁ ~ bm is a process of dispensing and arranging equal amounts of _{m m} reagents in m column × 1 row container groups, and the step (S2) is performed on all containers arranged in a matrix of n columns × m rows. Dispensing an amount of magnetic particles to bind the target substance in the reagent with the magnetic particles, and the step (S3) is performed in all containers arranged in a matrix of the n columns × m rows. In addition, a magnetic field is applied by a magnetic means provided outside the container, and a combined body with magnetic particles is adsorbed and held on the inner surface of each container, and other than the combined body adsorbed and held on the inner surface of the container by the pipette means The container of n columns × m rows or the pipette means nozzle group is removed by suction. By location moves, the entire container in which the conjugate are arranged in a matrix of the n columns × m rows contained, the reagent contained in each container of the b ₁ ~b _m and n equal parts The combinatorial synthesis method using magnetic particles according to claim 9, wherein the method is a process of dividing and transporting the particles. The step (S1) is in each n columns × m rows container group n columns × m rows of a matrix in arrayed m / n Gunko, the n-number of each type of reagent a ₁ ~ a _n, The step (S2) is a process of dispensing an equal amount into m container groups in each column, and the process (S2) includes n / m groups of n columns × m rows of container groups. A step of dispensing an equal amount of magnetic particles into all the containers of a row × m row container group to combine a target substance and magnetic particles contained in each reagent to form a conjugate, (S3) is a first group of n columns × m rows of container groups using pipette means in which magnetic means is provided only for the combined body among the contents of each container of each of the n columns × m rows of container groups. 10. The process of transferring from the first group to the next n-th column × m-th row container group by transposition or translational movement until reaching the m-th group n-th column × m-th row container group. Combinatorial synthesis method using magnetic particles according. Substances bonded to magnetic particles by adding an identifier for identifying each substance together with each substance of the substance group, and binding the identifier to the magnetic particles or positioning the identifier around the magnetic particles The combinatorial synthesis method using the magnetic particles according to any one of claims 9 to 12, wherein:

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