CN104231037A - Rapid protein purification combination chromatographic column, kit and preparation method - Google Patents
Rapid protein purification combination chromatographic column, kit and preparation method Download PDFInfo
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- CN104231037A CN104231037A CN201410474278.2A CN201410474278A CN104231037A CN 104231037 A CN104231037 A CN 104231037A CN 201410474278 A CN201410474278 A CN 201410474278A CN 104231037 A CN104231037 A CN 104231037A
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Abstract
The invention provides a rapid protein purification combination chromatographic column, a kit and a preparation method. The chromatographic column comprises a column body, an ion exchange resin layer, three filter discs, a gel filtration resin layer and an outlet, wherein the three filter discs are arranged on the bottom surface in the column body, the bed surface of the ion exchange resin layer and the bed surface of the gel filtration resin layer respectively, the shapes and the sizes of the filter discs are identical to those of the section of the column body, pore diameters are larger than the diameter of separated protein and smaller than diameters of ion exchange resin and gel filtration resin, and the outlet is formed in the bottom of the column body. The rapid protein purification combination chromatographic column is used for performing protein separation and purification and is convenient and rapid to operate, time can be greatly saved, and the working efficiency is improved.
Description
Technical field
The invention belongs to separation and purification of protein technical field, specifically relate to the combination of a kind of FPLC purifying chromatography column, test kit and preparation method.
Background technology
Protein separation is the method going out required target protein with biotechnology downstream technique from separation and purification among mixture, is an important technology of biological technical field.This technical difficulty, cost are all high, and such as 75% of a biologics cost all spends in the middle of downstream protein separation and purification.Protein separation technology mainly contains precipitation, dialysis, chromatography etc., and the common technology wherein in chromatographic technique has ion exchange chromatography and molecular sieve (gel-filtration).
Ion exchange chromatography (Ion Exchange Chromatography, IEC) being take ion-exchanger as stationary phase, a kind of chromatography method that the difference of bonding force size when carrying out reversible exchange according to the component ion in moving phase and the counterion on exchanger and carrying out is separated.1848, the people such as Thompson found ion-exchange phenomenon in research surface soil alkalinity material exchange process.1940's, there is the polystyrene ion-exchange resin with stable commutativity.The fifties, ion exchange chromatography enters biochemical field, is applied to amino acid whose analysis.At present, ion exchange chromatography is still a kind of chromatography method conventional in biochemical field, is widely used in the separation and purification of various biochemical substances as amino acid, albumen, carbohydrate, Nucleotide etc.According to the performance of ion exchange resin, be divided into Zeo-karb and anionite-exchange resin.If separated material is positively charged basic protein, they are more stable in an acidic solution, avidity is strong, therefore employing cationite, and acidic substance, more stable in basic solution, then use anionite, which kind of if be zwitter-ion for the material be separated, generally consider with electric charge in its stable pH value range, the selection of in return agent.Mainly processing power is large for the advantage of ion exchange resin, and separating ranges is wide, is separated capacity high, can removes various different ion, can use by repeated regeneration, long working life, working cost lower (although a Meteorological is larger).
Gel-filtration is the porousness utilizing gel (as dextrane gel, sepharose, polyacrylamide gel etc.), according to molecular size, the different speed difference be diffused in gel pore of shape of separated material, a kind of chromatography be thus separated by the speed difference of chromatography column.Gel chromatography has been widely used in the separating-purifying of the materials such as enzyme, protein, amino acid, polysaccharide, hormone, alkaloid.The advantage of gel-filtration mainly contains separation condition gentleness, and protein is volatility not, and yield is high, favorable reproducibility, and working range is wide, the wide coverage etc. of isolated molecule amount.
Because ion exchange resin is different from the separation principle of gel filtration resin, two kinds of chromatographies alternately use can be used for being separated, purification of high-purity albumen.But such separation and purification of protein process is a very loaded down with trivial details process.Usual protein crude extract is first added to after resinbed analyses post, through washing post, gradient elution, collect the component that different time flows out, protein crude extract is made to be separated into multiple component, each component also needs to analyze further through additive method (as SDS-PAGE gel electrophoresis) to find out containing target protein component, then be separated adding on gel filtration resin chromatography column containing target protein component further, collect the component that different time flows out, again through component that additive method (as SDS-PAGE gel electrophoresis) analysis is found out containing relative high purity target protein.Such separation and purification of protein process not only needs multiple chromatography column, also needs to design complicated buffer solution system, does not have professional training, cannot be engaged in such work.Therefore, a kind of simple FPLC separation and purification chromatography column of exploitation is necessary.
Summary of the invention
The problem to be solved in the present invention is to provide the chromatography column of a kind of conbined usage ion exchange resin and gel filtration resin, carries out albumen sepn and purifying with this chromatography column, easy to operate, quick, greatly can save time, increase work efficiency.
For solving the problems of the technologies described above, the technical solution used in the present invention is: a kind of FPLC purifying combination chromatography column, comprise cylinder, resinbed, three filter discs, gel filtration resin layer, spouts, described three filter discs are arranged on column body bottom surface, resinbed bed surface and on gel filtration resin layer bed surface respectively, filter disc shape size is identical with pole section, aperture is greater than separated albumen, be less than ion exchange resin and gel filtration resin, spout is arranged on bottom cylinder.
The cylinder of chromatography column is plastic column, glass column or metal column, and cylinder body shape is cylindrical, square column type or elliptical cylinder-shape, and filter disc is plastics filter disc or paper filter disc.
Further, the packing height of described resinbed is 0.3-3 times of cylinder internal diameter, and the packing height of described gel filtration resin layer is more than 2 times of described resinbed packing height.
Further, the packing height of described gel filtration resin layer be described ion exchange resin packing height 5-20 doubly between.
Prepare a method for above-mentioned FPLC purifying combination chromatography column, comprise the steps:
(1) choose chromatography column void column body, after clean, bottom cylinder, put into a slice filter disc;
(2) gel filtration resin or the ion exchange resin of activation is got, with water-soluble swollen fully after, take a morsel loading cylinder, flows out by water by gravity settling, repeatedly loads resin, sedimentation, during as added gel filtration resin, bed surface is pressed into a slice filter disc gently, during as added ion exchange resin after reaching more than 0.6 times of cylinder internal diameter on bed surface, bed surface is pressed into a slice filter disc gently after reaching 0.3-3 times of cylinder internal diameter on bed surface;
(3) gel filtration resin or the ion exchange resin of activation is got, and it is not identical with the resin of step (2), with water-soluble swollen fully after, take a morsel above being added on step (2) resin layer bed surface filter disc, flow out by water by gravity settling, repeatedly load resin, sedimentation, during as added gel filtration resin, after bed surface reaches more than 2 times of the ion exchange resin bed face height that step (2) adds, bed surface is pressed into a slice filter disc gently, during as added ion exchange resin, the ratio of the gel filtration resin layer bed surface height added when bed surface height and step (2) is greater than 0, after being less than or equal to 0.5, bed surface is pressed into a slice filter disc gently,
(4) with 30% aqueous ethanolic solution of 50ml, make an addition on chromatography column by amount, open chromatography column lower exit port, be repeatedly added into after whole 30% aqueous ethanolic solution adds, cover post mouth above chromatography column lower exit port mouth and chromatography column by hd respectively, it is for subsequent use to be placed in refrigerator 4-8 DEG C of refrigeration.
A single stage method interleukin purification kit containing above-mentioned FPLC purifying combination chromatography column, also comprises damping fluid.
Further, described damping fluid comprises one or both in Glycine-NaOH damping fluid and Tris-HCl damping fluid.
A using method for single stage method interleukin purification kit, comprises the steps:
(1) get damping fluid deionized water and be diluted to 1X damping fluid;
(2) 5 times are combined chromatography column ion exchange resin and the overall accumulated amount of gel filtration resin 1X damping fluid to FPLC purifying is got, above post, FPLC purifying combination chromatography column is made an addition to by amount, make it by chromatography column, with resin buffer system in balance columns;
(3) by this 1X damping fluid dialysed overnight of protein crude extract to be separated, or in protein crude extract to be separated, add appropriate concentrated damping fluid, the final concentration of damping fluid is made to be 1X, whether the protein crude extract pH value detected after dilution is identical with the pH value of former damping fluid, regulates its pH value to identical with former pH of cushioning fluid;
(4) protein crude extract that the 1/10-1/20 getting gel filtration resin volume processes through step (3), be added on the upper strata resin of the FPLC purifying combination chromatography column processed through step (2), protein crude extract is allowed to flow in resin by deadweight or pressure difference, then 3-5 doubly combines the overall accumulated amount of resin in chromatography column 1X damping fluid to FPLC purifying is got, it is allowed to pass through chromatography column, make the separated rear outflow of target protein, eluting peak is detected at 280nm wavelength place with Ultraviolet Detector, or detection signal is inputted chromatographic working station system, draw elution curve,
(5) from albumen outflow component being detected, component collects effluent liquid component, detects target protein purity and content in each effluent liquid component, preserves the effluent liquid component containing target protein.
The advantage that the present invention has and positively effect are:
1, the present invention adopts and ion exchange resin and gel filtration resin layering is put into same chromatography column, centre filter disc separates, one time upper prop can carry out ion exchange chromatography and gel permeation chromatography simultaneously, without the need to washing and the gradient elution supervisor of complexity, without the need to using multiple chromatography column, simple operation, routine experimentation personnel get final product skilled operation after simple training, save time, save reagent consumptive material;
2, all purge process only needs a kind of damping fluid, without the need to designing the damping fluid of multiple gradient salinity damping fluid and different pH value, making simple, convenient, increasing work efficiency;
3, the interleukin purification kit comprising FPLC purifying combination chromatography column of the present invention, single stage method fast separating and purifying interleukin albumen, obtain highly purified target protein, easy to use, reagent cost is low.
Accompanying drawing explanation
Fig. 1 is I-type FPLC purifying of the present invention combination chromatography column.
Fig. 2 is II-type FPLC purifying of the present invention combination chromatography column.
Fig. 3 is target protein IL-4 purity in each component of SDS-PAGE electrophoresis detection of the present invention.
Fig. 4 is target protein IL-7 purity in each component of SDS-PAGE electrophoresis detection of the present invention.
In Fig. 1 and Fig. 2: 1. cylinder, 2. resinbed, 3. filter disc, 4. gel filtration resin layer, 5. spout.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described, but do not limit protection scope of the present invention.
Embodiment 1
As shown in Figure 1, a kind of FPLC purifying combination chromatography column (I-type), plastics filter disc 3, Sephadex G-50 gel filtration resin layer 4, plastics filter disc 3, DEAE anion exchange resin layer 2, plastics filter disc 3 from bottom to top respectively in the round plastic chromatographic column cylinder 1 of diameter 20mm, long 300mm, the shape size of plastics filter disc 3 is identical with cylinder 1 cross section, aperture is greater than separated albumen, is less than DEAE anionite-exchange resin and Sephadex G-50 gel filtration resin.
One prepares the method for FPLC purifying combination chromatography column (I-type), comprises the steps:
(1) choose the round plastic chromatography column void column body 1 of a diameter 20mm, long 300mm, bottom cylinder 1, after clean, put into the plastics filter disc 3 that a slice is equal to cylinder 1 internal diameter;
(2) the Sephadex G-50 gel filtration resin of 10g activation is got, with water-soluble swollen fully after, take a morsel and load cylinder 1, flow out by water by gravity settling, repeatedly load resin, sedimentation, until the height of bed of gel filtration resin layer 4 reaches 200mm after sedimentation, residue Sephadex G-50 gel filtration resin does not reinstall, and this bed surface is pressed into plastics filter disc 3 that a slice is equal to cylinder 1 internal diameter gently;
(3) the DEAE anionite-exchange resin of 2g activation is got, with water-soluble swollen fully after, take a morsel above being added on gel filtration resin layer 4 bed surface filter disc 3, flow out by water by gravity settling, repeatedly load resin, sedimentation, until the height of bed of resin layer reaches 30mm after sedimentation, residue DEAE anionite-exchange resin does not reinstall, and this bed surface is pressed into plastics filter disc 3 that a slice is equal to cylinder 1 internal diameter more gently;
(4) chromatography column is prepared complete, if used immediately, uses after available buffer liquid balance pillar; If do not used immediately, cover post mouth above chromatography column lower exit port mouth and chromatography column by hd respectively after available 30% aqueous ethanolic solution balance pillar, it is for subsequent use to be placed in refrigerator 4-8 DEG C of refrigeration.
Embodiment 2
As shown in Figure 2, a kind of FPLC purifying combination chromatography column (II-type), plastics filter disc 3, DEAE anion exchange resin layer 2, plastics filter disc 3, Sephadex G-50 gel filtration resin layer 4, plastics filter disc 3 from bottom to top respectively in the round plastic chromatographic column cylinder 1 of diameter 20mm, long 300mm, the shape size of plastics filter disc 3 is identical with cylinder 1 cross section, aperture is greater than separated albumen, is less than DEAE anionite-exchange resin 2 and Sephadex G-50 gel filtration resin 4.
One prepares the method for FPLC purifying combination chromatography column (II-type), comprises the steps:
(1) choose the round plastic chromatography column void column body 1 of a diameter 20mm, long 300mm, at the bottom of post, after clean, put into the plastics filter disc 3 that a slice is equal to cylinder 1 internal diameter;
(2) the DEAE anionite-exchange resin of 2g activation is got, with water-soluble swollen fully after, take a morsel and load cylinder 1, flow out by water by gravity settling, repeatedly load resin, sedimentation, until the height of bed of DEAE anion exchange resin layer 2 reaches 25mm after sedimentation, residue DEAE anionite-exchange resin does not reinstall, and this bed surface is pressed into a slice more gently and is equal to plastics filter disc 3 with cylinder 1 internal diameter;
(3) the Sephadex G-50 gel filtration resin of 10g activation is got, with water-soluble swollen fully after, take a morsel above being added on DEAE anion exchange resin bed face filter disc 3, flow out by water by gravity settling, repeatedly load resin, sedimentation, reach 200mm from the height of bed of gel filtration resin layer 4 to sedimentation, residue Sephadex G-50 gel filtration resin does not reinstall, and this bed surface is pressed into plastics filter disc 3 that a slice is equal to cylinder 1 internal diameter gently;
(4) chromatography column is prepared complete, if used immediately, uses after available buffer liquid balance pillar; If do not used immediately, after available 30% aqueous ethanolic solution balance pillar, cover post mouth above chromatography column lower exit port mouth and chromatography column by hd respectively, it is for subsequent use to be placed in refrigerator 4-8 DEG C of refrigeration.
Embodiment 3
A single stage method interleukin purification kit containing FPLC purifying combination chromatography column, comprises I-type FPLC purifying combination chromatography column, one bottle of 20X Glycine-NaOH and concentrates damping fluid.
A preparation method for single stage method interleukin purification kit, comprises the steps:
(1) choose a diameter 10mm, the round plastic chromatography column void column body 1 of long 350mm, at the bottom of post, put into a slice after clean and be equal to plastics filter disc 3 with cylinder 1 internal diameter;
(2) the Sephadex G-50 gel filtration resin 4 of 10g activation is got, with water-soluble swollen fully after, load cylinder 1, flow out by water by gravity settling, repeatedly load resin, sedimentation, until the height of bed of gel filtration resin layer 4 reaches 250mm after sedimentation, residue Sephadex G-50 gel filtration resin does not reinstall, and this bed surface is pressed into a slice gently and is equal to plastics filter disc 3 with cylinder 1 internal diameter;
(3) the CM weak cation exchange resin of 1g activation is got, with water-soluble swollen fully after, take a morsel above being added on gel filtration resin layer 4 bed surface filter disc 3, flow out by water by gravity settling, repeatedly load resin, sedimentation, until the height of bed of CM weak cation exchange resin layer 2 reaches 20mm after sedimentation, residue weak cation exchange resin does not reinstall, and this bed surface is pressed into a slice more gently and is equal to plastics filter disc 3 with cylinder 1 internal diameter.
(4) 50ml30% aqueous ethanolic solution is configured, it is made an addition on above-mentioned prepared chromatography column by amount, open chromatography column lower exit port, repeatedly be added into after whole 30% aqueous ethanolic solution adds, cover chromatography column lower exit port mouth by hd respectively and analyse post mouth above post, it is for subsequent use to be placed in refrigerator 4-8 DEG C of refrigeration.
(5) prepare the 500mM Glycine-NaOH damping fluid of the pH=10 that 20X concentrates, comprise the steps:
1) glycine (molecular weight: 70.07) 35g is taken, add deionized water to 1L, stir evenly and make fully to dissolve just to obtain 500mM glycine solution, weighing sodium hydroxide (molecular weight: 40) 20g, add deionized water to 1L, stir evenly and make fully to dissolve just to obtain 0.5N sodium hydroxide solution;
2) 500mM glycine solution 500ml is got, get 0.5N sodium hydroxide solution 320ml, stirring and in 500mM glycine solution, progressively adding 0.5N sodium hydroxide solution till pH=10 under measuring pH value situation, just the Glycine-NaOH damping fluid of the pH=10 that 20X concentrates is obtained, load in Plastic Bottle, it is for subsequent use to be placed in refrigerator 4-8 DEG C of refrigeration.
(6) the 20X Glycine-NaOH getting I-type FPLC purifying combination chromatography column one and the one bottle 500ml of above-mentioned preparation concentrates damping fluid, attaches together in a paper box, obtains single stage method interleukin purification kit.
Embodiment 4
The using method of single stage method interleukin Purification Kit recombination human interleukin-4 (rhIL-4, molecular weight 14.8KD, iso-electric point 9.3), comprises the steps:
(1) get single stage method interleukin purification kit a set of, comprise I-type FPLC purifying combination chromatography column and one bottle of 20X concentrates Glycine-NaOH damping fluid;
(2) from test kit, get 20X and concentrate damping fluid 50ml, be diluted to 1X damping fluid 1000ml with deionized water;
(3) from test kit, take out I-type FPLC purifying combination chromatography column, open above chromatography column and here lid, get 1X damping fluid 100ml, above post, make an addition to FPLC purifying combination chromatography column by amount, make it by chromatography column, with wash and balance columns in resin buffer system;
(4) by the 1X damping fluid dialysed overnight of the crude extract 2ml 500ml of the escherichia coli expression containing rhIL-4;
(5) protein crude extract of will dialyse, be added on the upper strata resin of equilibrated I-type FPLC purifying combination chromatography column, protein crude extract is allowed to flow in resin by deadweight, then the 1X damping fluid of 50ml is got, add chromatography column by amount, make the separated rear outflow of target protein, detect eluting peak with Ultraviolet Detector at 280nm wavelength place, or detection signal is inputted chromatographic working station system, draw elution curve;
(6) from detecting that albumen collects effluent liquid component, with target protein purity and content in each component of SDS-PAGE electrophoresis detection by every 1ml component flowing out component;
(7) analyze according to SDS-PAGE electrophorogram, as shown in Figure 3, effluent liquid component 3,4 (namely the 4th, 5 swimming lanes) has the highest target protein IL-4 of purity, preserves this component.
Embodiment 5
The using method of single stage method interleukin Purification Kit recombination human interleukin-7 (rhIL-7, molecular weight 17.2KD, iso-electric point 8.9), comprises the steps:
(1) get single stage method interleukin purification kit a set of, comprise the 20mM Tris-HCl damping fluid 500ml of an II-type FPLC purifying combination chromatography column and one bottle of PH=8;
(2) open above II-type FPLC purifying combination chromatography column and here lid, get Tris-HCl damping fluid and be about 100ml, by amount make an addition to above post FPLC purifying combination chromatography column, make it by chromatography column, with wash and balance columns in resin buffer system;
(3) by the crude extract 3ml 500mlTris-HCl damping fluid dialysed overnight of the escherichia coli expression containing rhIL-7, by the protein crude extract of dialysing, be added on the upper strata resin of equilibrated II-type FPLC purifying combination chromatography column, protein crude extract is allowed to flow in resin by deadweight, then the 1X damping fluid of 50ml is got, chromatography column is added by amount, make the separated outflow of target protein, eluting peak is detected at 280nm place with Ultraviolet Detector, or detection signal is inputted chromatographic working station system, draw elution curve;
(4) from detecting that albumen collects effluent liquid component, with target protein purity and content in each component of SDS-PAGE electrophoresis detection by every 2ml component flowing out component;
(5) analyze according to SDS-PAGE electrophorogram, as shown in Figure 4, effluent liquid component 6 (i.e. the 6th swimming lane) has the highest target protein IL-7 of purity, preserves this component.
Above preferred embodiment of the present invention has been described in detail, but described content being only preferred embodiment of the present invention, can not being considered to for limiting practical range of the present invention.All equalizations done according to the present patent application scope change and improve, and all should still belong within patent covering scope of the present invention.
Claims (7)
1. a FPLC purifying combination chromatography column, it is characterized in that: comprise cylinder, resinbed, three filter discs, gel filtration resin layer, spouts, described three filter discs are arranged on column body bottom surface, resinbed bed surface and on gel filtration resin layer bed surface respectively, filter disc shape size is identical with pole section, aperture is greater than separated albumen, be less than ion exchange resin and gel filtration resin, spout is arranged on bottom cylinder.
2. a kind of FPLC purifying combination chromatography column according to claim 1, it is characterized in that: the packing height of described resinbed is 0.3-3 times of cylinder internal diameter, and the packing height of described gel filtration resin layer is more than 2 times of described resinbed packing height.
3. a kind of FPLC purifying combination chromatography column according to claim 2, is characterized in that: the packing height of described gel filtration resin layer be described resinbed packing height 5-20 doubly between.
4. prepare a method for chromatography column described in any one of claim 1-3, it is characterized in that: comprise the steps:
(1) choose chromatography column void column body, after clean, bottom cylinder, put into a slice filter disc;
(2) gel filtration resin or the ion exchange resin of activation is got, with water-soluble swollen fully after, take a morsel loading cylinder, flows out by water by gravity settling, repeatedly loads resin, sedimentation, during as added gel filtration resin, bed surface is pressed into a slice filter disc gently, during as added ion exchange resin after reaching more than 0.6 times of cylinder internal diameter on bed surface, bed surface is pressed into a slice filter disc gently after reaching 0.3-3 times of cylinder internal diameter on bed surface;
(3) gel filtration resin or the ion exchange resin of activation is got, and it is not identical with the resin of step (2), with water-soluble swollen fully after, take a morsel above being added on step (2) resin layer bed surface filter disc, flow out by water by gravity settling, repeatedly load resin, sedimentation, during as added gel filtration resin, after bed surface reaches more than 2 times of the ion exchange resin bed face height that step (2) adds, bed surface is pressed into a slice filter disc gently, during as added ion exchange resin, the ratio of the gel filtration resin layer bed surface height added when bed surface height and step (2) is greater than 0, after being less than or equal to 0.5, bed surface is pressed into a slice filter disc gently,
(4) with 30% aqueous ethanolic solution of 50ml, make an addition on chromatography column by amount, open chromatography column lower exit port, be repeatedly added into after whole 30% aqueous ethanolic solution adds, cover post mouth above chromatography column lower exit port mouth and chromatography column by hd respectively, it is for subsequent use to be placed in refrigerator 4-8 DEG C of refrigeration.
5. the single stage method interleukin purification kit containing chromatography column described in any one of claim 1-3, is characterized in that: also comprise damping fluid.
6. a kind of single stage method interleukin purification kit according to claim 5, is characterized in that: described damping fluid comprise in Glycine-NaOH damping fluid and Tris-HCl damping fluid one or both.
7. a using method for test kit described in claim 5 or 6, is characterized in that: comprise the steps:
(1) get damping fluid deionized water and be diluted to 1X damping fluid;
(2) 5 times are combined chromatography column ion exchange resin and the overall accumulated amount of gel filtration resin 1X damping fluid to FPLC purifying is got, above post, FPLC purifying combination chromatography column is made an addition to by amount, make it by chromatography column, with resin buffer system in balance columns;
(3) by this 1X damping fluid dialysed overnight of protein crude extract to be separated, or in protein crude extract to be separated, add appropriate concentrated damping fluid, the final concentration of damping fluid is made to be 1X, whether the protein crude extract pH value detected after dilution is identical with the pH value of former damping fluid, regulates its pH value to identical with former pH of cushioning fluid;
(4) protein crude extract that the 1/10-1/20 getting gel filtration resin volume processes through step (3), be added on the upper strata resin of the FPLC purifying combination chromatography column processed through step (2), protein crude extract is allowed to flow in resin by deadweight or pressure difference, then 3-5 doubly combines the overall accumulated amount of resin in chromatography column 1X damping fluid to FPLC purifying is got, it is allowed to pass through chromatography column, make the separated rear outflow of target protein, eluting peak is detected at 280nm wavelength place with Ultraviolet Detector, or detection signal is inputted chromatographic working station system, draw elution curve,
(5) from albumen outflow component being detected, component collects effluent liquid component, detects target protein purity and content in each effluent liquid component, preserves the effluent liquid component containing target protein.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2020030099A1 (en) * | 2018-08-10 | 2020-02-13 | 浙江楚沅生物科技有限公司 | Method for purifying active substances in non-human animal amniotic fluid |
| WO2023060622A1 (en) * | 2021-10-17 | 2023-04-20 | 南通市台盈新材料科技有限公司 | Enrichment tube for enriching total flavones in epimedium brevicomu maxim leaf extracting solution |
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2014
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2020030099A1 (en) * | 2018-08-10 | 2020-02-13 | 浙江楚沅生物科技有限公司 | Method for purifying active substances in non-human animal amniotic fluid |
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| CN110818773B (en) * | 2018-08-10 | 2023-04-11 | 浙江楚沅生物科技有限公司 | Method for purifying active substance in amniotic fluid of non-human animal |
| WO2023060622A1 (en) * | 2021-10-17 | 2023-04-20 | 南通市台盈新材料科技有限公司 | Enrichment tube for enriching total flavones in epimedium brevicomu maxim leaf extracting solution |
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