JPS62235568A - Carrier biochemically activated by antibody covalent-bonded with surface - Google Patents
Carrier biochemically activated by antibody covalent-bonded with surfaceInfo
- Publication number
- JPS62235568A JPS62235568A JP61290299A JP29029986A JPS62235568A JP S62235568 A JPS62235568 A JP S62235568A JP 61290299 A JP61290299 A JP 61290299A JP 29029986 A JP29029986 A JP 29029986A JP S62235568 A JPS62235568 A JP S62235568A
- Authority
- JP
- Japan
- Prior art keywords
- carrier
- antibody
- peptide chain
- binding site
- complementary
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000009739 binding Methods 0.000 claims description 12
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 10
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 7
- 239000000126 substance Substances 0.000 claims description 7
- 230000000295 complement effect Effects 0.000 claims description 6
- 239000012634 fragment Substances 0.000 claims description 5
- 210000004027 cell Anatomy 0.000 claims description 3
- 125000000466 oxiranyl group Chemical group 0.000 claims description 3
- 230000035755 proliferation Effects 0.000 claims description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 claims 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 claims 1
- 210000000988 bone and bone Anatomy 0.000 claims 1
- 210000000845 cartilage Anatomy 0.000 claims 1
- 210000000170 cell membrane Anatomy 0.000 claims 1
- 230000004069 differentiation Effects 0.000 claims 1
- 210000002744 extracellular matrix Anatomy 0.000 claims 1
- 210000000056 organ Anatomy 0.000 claims 1
- 239000007943 implant Substances 0.000 description 9
- 102000007350 Bone Morphogenetic Proteins Human genes 0.000 description 7
- 108010007726 Bone Morphogenetic Proteins Proteins 0.000 description 7
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 7
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 7
- 229940112869 bone morphogenetic protein Drugs 0.000 description 7
- 229920001577 copolymer Polymers 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical group C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 230000000274 adsorptive effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000004821 effect on bone Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- -1 proteins Chemical class 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 238000007493 shaping process Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
Landscapes
- Materials For Medical Uses (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【産業上の利用分野１
本発明は、表面に共有結合した抗体によって生化学的に
活性化された担体に関するものであり、本発明担体は体
内移植体として使用するのに好適である。
【従来の技術】
生化学的活性化合物、例えば蛋白質、ｉＰ！素、抗体等
を、変性した担鉢物買上に固定すること、そして医学、
薬学、バイオテクノロジー、化学、ＩＮ！業等に利用す
ることは知られており、例えばドイツ特許明１［１占用
２１０６９３号に記載されている。例えば、共有結合に
よって担体に固定された抗体は、例えば体液中にあるホ
ルモンのような物質を特定するための分析反応を取扱う
のに広く応用される（ドイツ特許明［１第２２０６１０
３号参照）。更にフランス特許明１１１１！第２５５４
３４９号には、担体に共有結合した特異的モノクローナ
ル抗体よって、相応する抗凝血物質、例えばヘパリンを
結合させることが記載されている。このように形成され
た担体は人工器官として用いることができる。
Ｋ発明が解決しようとする問題点】
本発明の目的は、これとは異なり、活発な細胞集団増殖
を可能にするような表面の性質を有する担体を提供する
ことである。更に本発明は担体の表面の性質を、人体に
ある物質によって誘導され得る活発な細胞集団増殖が可
能であるように形成することを目的とする。[Industrial Application Field 1] The present invention relates to a carrier biochemically activated by an antibody covalently bonded to the surface, and the carrier of the present invention is suitable for use as an implant in the body. [Prior Art] Biochemically active compounds, such as proteins, iP! Immobilizing antibodies, antibodies, etc. on denatured potable materials, and medicine,
Pharmacy, biotechnology, chemistry, IN! It is known that it can be used in industrial applications, and is described, for example, in German Patent No. 1 [1 Utsuyō 210693]. For example, antibodies immobilized by covalent bonds to carriers are widely applied in handling analytical reactions, for example to identify substances such as hormones in body fluids (German Patent No. 1 220 610
(See No. 3). Furthermore, French patent Ming 1111! No. 2554
No. 349 describes the binding of corresponding anticoagulants, such as heparin, by means of specific monoclonal antibodies covalently bound to carriers. A carrier formed in this way can be used as a prosthesis. K Problems to be Solved by the Invention It is an object of the present invention to provide a carrier having surface properties that allow active cell population growth. Furthermore, the present invention aims at shaping the surface properties of the carrier in such a way that active cell population growth, which can be induced by substances present in the human body, is possible.
上述の目的は、本発明により、表面に共有結合された抗
体によって生化学的に活性化された担体によっｔ達せら
れる。即ち、細胞集団の増殖及び／又は分化を調節する
ためには、担体表面に結合する抗体又は抗体切断片又は
抗体切断片部分のペプチド鎖の結合部位が、人体内にお
いて自然に生成される物質である蛋白質の決定基に相補
的であることである。
高度に特異的なモノクローナル抗体を、現在利用できる
技術によって、殆ど全ての所望のエピトープ（抗原決定
基）に対して作り出すことができる。モノクローナル抗
体を作るためには極く少量の抗体量で十分である。人体
固有の多数の物質とは異なり、モノクローナル抗体は、
［ｌ菌又は哺乳類細胞の遺伝子工学的変形を必要とする
ことなく、キログラム吊で経済的に生産することができ
る。
モノクローナル抗体又はその切断片を、再生可能に固体
表面に共有結合させる。抗体を、例えばパパインによっ
て蛋白分解的に分離し、それから例えば電気泳動又はク
ロマトグラフィーによって精製した後、Ｎ−末端に結合
部位を有するペプチド鎖、特にペプチド鎖の変・わり得
る部分を担体表面に共有結合させる。共有結合は、担体
表面にあるオキシラン基を介して行われる。このために
担体表面は適当な重合物質によって形成される。その他
の好適な共有結合の種類はケミカー−ツ？イトウング（
Ｃｈｅｍｉｋｅｒ−２ｅｉｔｕｎＱ）９７版（１９７３
）ＮＯ，１１，１３１２頁に記載されている。
本発明による抗体或いは抗体切断片或いは抗体切断片部
分のペプチド鎖の末端或いは結合部位の形成によって、
体内にある蛋白質のためのこれらレセプターが形成され
る。このような担体表面に結合したモノクローナル抗体
、或いは抗体切断片或いは該切断片部分は安定で、長期
間人体内にあっても酵素分解を受けない。その特異性の
ために、人体固有の分子が分子の活性中心が成長に最適
の状態にあるように一定の方法で、エピトープで担体表
面に吸着されることが可能となる。担体表面としては特
に移植体表面が問題となる。モノクローナル抗体で習わ
れた担体表面上に人体固有の物質が吸着的に結合するこ
とによって、移植体表面が人体固有の物質で効果的に覆
われることになる。
特に、モノクローナル抗体は骨形成蛋白質（Ｂｏｎｅ
Ｈｏｒｐｈｏｇｅｎｉｃ ｐｒｏｔａｉｎｓ）に対し結
合能力を有するため、移植後、移植体表面にある抗体或
いは抗体切断片或いは抗体切断片部分が患者の骨形成蛋
白質を吸着結合して、安定な生化学的に活性な移植体表
面が得られる。
に実 浦 例１
本発明を説明するために、次に、実施例として移植体の
製造について記す。
実施例
移植体の金属核を、少なくとも表面にオキシラン基を有
し、それによって抗体が共有結合できる共重合体で被覆
する。抗体を担体表面に結合させた後、共重合体表面か
らまだ残っているモノマーを抽出する。結合反応の前に
、場合によってはモノクローナル抗体を蛋白分解によっ
て切り離し、変化し得る部分或いは切り離されたペプチ
ド鎖を活性化された共重合体表面に共有結合させる。こ
の共有結合はオキシラン基によって行われる・移植後、
移植体は骨形成蛋白質によって被覆され、その際骨形成
蛋白質は一定の部位（エピトープ）で抗体に吸着される
。この方法で骨形成蛋白質の移植体表面への固定が達成
される。
Ｋ発明の効果１
この本発明の方法で臂られた移植体表面は蛋白分解酵素
による分解に対して高い安定性を有する。
骨形成蛋白質は吸着的に結合する。万一蛋白分解酵素に
よって蛋白質の分解がおきた場合には、特にその分解に
よっておきる構造上の変化に基づいて、そこなわれてい
ない完全な骨形成蛋白質がとって代る。抗体に吸着さ゛
れた蛋白質はその活性を完全に保持するから、移植体挿
入後の骨形成において充分な効果を発揮することができ
る。The above-mentioned object is achieved according to the invention by means of a biochemically activated carrier with antibodies covalently bound to its surface. That is, in order to regulate the proliferation and/or differentiation of cell populations, the binding site of the peptide chain of the antibody or antibody fragment or antibody fragment portion that binds to the carrier surface must be a substance naturally produced in the human body. It is complementary to a determinant of a protein. Highly specific monoclonal antibodies can be generated against almost any desired epitope (antigenic determinant) using currently available technology. A very small amount of antibody is sufficient to produce monoclonal antibodies. Unlike many substances unique to the human body, monoclonal antibodies
It can be economically produced in kilogram quantities without the need for genetic engineering modification of bacteria or mammalian cells. A monoclonal antibody or truncated fragment thereof is reproducibly covalently attached to a solid surface. After the antibody has been proteolytically separated, e.g. by papain, and then purified, e.g. by electrophoresis or chromatography, the peptide chain, in particular the variable part of the peptide chain, with the binding site at the N-terminus is covalently attached to the surface of the carrier. combine. Covalent bonding takes place via oxirane groups on the surface of the carrier. For this purpose, the carrier surface is formed by a suitable polymeric material. What are other suitable types of covalent bonds? Itung (
Chemiker-2eitunQ) 97th edition (1973
) No. 11, page 1312. By forming the terminal or binding site of the peptide chain of the antibody, antibody fragment, or antibody fragment portion according to the present invention,
These receptors for proteins found in the body are formed. The monoclonal antibody, the antibody fragment, or the fragment portion bound to the surface of such a carrier is stable and does not undergo enzymatic degradation even if it remains in the human body for a long period of time. Its specificity allows molecules specific to the human body to be epitopically adsorbed onto the carrier surface in a certain way such that the active center of the molecule is in an optimal state for growth. The surface of the carrier is particularly problematic, especially the surface of the implant. Adsorptive binding of human body-specific substances onto the carrier surface, as learned from monoclonal antibodies, effectively covers the implant surface with human body-specific substances. In particular, monoclonal antibodies target bone morphogenetic proteins (Bone morphogenetic proteins).
After transplantation, the antibody or antibody fragment or antibody fragment portion on the surface of the implant adsorbs and binds the patient's bone morphogenetic proteins, resulting in a stable biochemically active implant. The body surface is obtained. Example 1 In order to explain the present invention, production of an implant will now be described as an example. EXAMPLES The metal core of the implant is coated with a copolymer having oxirane groups at least on its surface to which antibodies can be covalently attached. After binding the antibody to the carrier surface, any remaining monomers are extracted from the copolymer surface. Prior to the binding reaction, the monoclonal antibody is optionally proteolytically cleaved and the variable moiety or cleaved peptide chain covalently attached to the activated copolymer surface. This covalent bond is carried out by the oxirane group・After grafting,
The implant is coated with bone morphogenetic proteins, which are then adsorbed to antibodies at specific sites (epitopes). In this way, fixation of bone morphogenetic proteins to the implant surface is achieved. K Effect of the Invention 1 The surface of the graft prepared by the method of the present invention has high stability against degradation by proteolytic enzymes. Bone morphogenetic proteins bind adsorbively. If a protein is degraded by a proteolytic enzyme, it will be replaced by an intact bone morphogenetic protein, especially on the basis of the structural changes caused by the degradation. Since the protein adsorbed to the antibody completely retains its activity, it can exert a sufficient effect on bone formation after implantation.
Claims (1)
された担体であって、細胞集団の増殖、分化を調節する
ために、担体表面に結合した抗体、抗体切断片、抗体切
断片部分の何れかのペプチド鎖の結合部位が、人体内で
自然に生成される物質を形成する蛋白質の決定基に対し
て相補的であることを特徴とする担体。 ２、抗体がモノクローナル抗体であることを特徴とする
特許請求の範囲第１項記載の担体。 ３、ペプチド鎖の結合部位が骨及び軟骨形成蛋白質に相
補的である特許請求の範囲第１又は第２項記載の担体。 ４、ペプチド鎖の結合部位が細胞外基質に相補的である
特許請求の範囲第１又は第２項記載の担体。 ５、ペプチド鎖の結合部位が免疫モジュレーターに相補
的であることを特徴とする特許請求の範囲第１又は第２
項記載の担体。 ６、ペプチド鎖の結合部位が細胞膜のエピトープに相補
的であることを特徴とする特許請求の範囲第１又は第２
項記載の担体。 ７、担体表面が、移植体又は臓器代替物の表面であるこ
とを特徴とする特許請求の範囲第１乃至第６項の中の何
れかの項に記載の担体。 ８、担体表面が、抗体或いはその断片と共有結合するた
めのオキシラン基を含むことを特徴とする特許請求の範
囲第１乃至第７項の何れかの項に記載の担体。[Scope of Claims] 1. A carrier biochemically activated by an antibody covalently bonded to the surface, the antibody or antibody bonded to the carrier surface in order to regulate the proliferation and differentiation of a cell population. A carrier characterized in that the binding site of the peptide chain of either the cut fragment or the antibody cut fragment is complementary to a determinant of a protein forming a substance naturally produced in the human body. 2. The carrier according to claim 1, wherein the antibody is a monoclonal antibody. 3. The carrier according to claim 1 or 2, wherein the binding site of the peptide chain is complementary to bone and cartilage forming proteins. 4. The carrier according to claim 1 or 2, wherein the binding site of the peptide chain is complementary to the extracellular matrix. 5. Claim 1 or 2, characterized in that the binding site of the peptide chain is complementary to the immune modulator
Carrier described in Section. 6. Claim 1 or 2, characterized in that the binding site of the peptide chain is complementary to an epitope of a cell membrane.
Carrier described in Section. 7. The carrier according to any one of claims 1 to 6, wherein the carrier surface is a surface of a transplant or an organ substitute. 8. The carrier according to any one of claims 1 to 7, wherein the carrier surface contains an oxirane group for covalent bonding with an antibody or a fragment thereof.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE3543066.4 | 1985-12-05 | ||
| DE3543066 | 1985-12-05 | ||
| DE3612643.8 | 1986-04-15 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS62235568A true JPS62235568A (en) | 1987-10-15 |
Family
ID=6287747
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61290299A Pending JPS62235568A (en) | 1985-12-05 | 1986-12-05 | Carrier biochemically activated by antibody covalent-bonded with surface |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62235568A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003526477A (en) * | 2000-03-15 | 2003-09-09 | オーバス メディカル テクノロジーズ インク． | Coating that promotes endothelial cell adhesion |
| US9320829B2 (en) | 2000-03-15 | 2016-04-26 | Orbusneich Medical, Inc. | Progenitor endothelial cell capturing with a drug eluting implantable medical device |
-
1986
- 1986-12-05 JP JP61290299A patent/JPS62235568A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003526477A (en) * | 2000-03-15 | 2003-09-09 | オーバス メディカル テクノロジーズ インク． | Coating that promotes endothelial cell adhesion |
| JP2013046771A (en) * | 2000-03-15 | 2013-03-07 | Orbusneich Medical Inc | Coating that promotes endothelial cell adherence |
| JP2015128598A (en) * | 2000-03-15 | 2015-07-16 | オーバスネイチ メディカル、インコーポレイテッド | Coating for promoting endothelial cell adhesion |
| US9320829B2 (en) | 2000-03-15 | 2016-04-26 | Orbusneich Medical, Inc. | Progenitor endothelial cell capturing with a drug eluting implantable medical device |
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