CN109336967A - Antibody purification process based on mixed fillers - Google Patents
Antibody purification process based on mixed fillers Download PDFInfo
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- CN109336967A CN109336967A CN201811332080.5A CN201811332080A CN109336967A CN 109336967 A CN109336967 A CN 109336967A CN 201811332080 A CN201811332080 A CN 201811332080A CN 109336967 A CN109336967 A CN 109336967A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/165—Extraction; Separation; Purification by chromatography mixed-mode chromatography
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/18—Ion-exchange chromatography
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/20—Partition-, reverse-phase or hydrophobic interaction chromatography
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Abstract
The present invention relates to protein stripping technique fields, more particularly, to a kind of antibody purification process based on mixed fillers.The antibody purification process based on mixed fillers includes the following steps: that chromatographing column using mixed fillers purifies crude product containing target antibody, and mixed fillers include anion-exchange chromatography medium and hydrophobic chromatoghaphy medium.Antibody purification process of the invention, by being used in mixed way anion-exchange chromatography medium and hydrophobic chromatoghaphy medium to obtain mixed fillers, consummate processing is carried out to antibody using mixed fillers one-step method, a variety of interactions can be generated with albumen, make full use of that mixed fillers adsorption capacity is big, good selective, separative efficiency is high, simplify separating step, reduce separation costs, the target antibody rate of recovery is high, the impurity such as high polymer can be effectively removed, solve the problems, such as to cannot be considered in terms of target antibody purity and the rate of recovery in the prior art.
Description
Technical field
The present invention relates to protein stripping technique fields, more particularly, to a kind of antibody purification side based on mixed fillers
Method.
Background technique
Antibody be can an albuminoid in conjunction with antigentic specificity, take part in the immunologic process of body, medicine, food,
The fields such as environment and agricultural have important application.Especially field of medicaments, monoclonal antibody strong, therapeutic effect with specificity
The advantages that significant and toxic side effect is small, is widely used in the diagnosing and treating of the major diseases such as autoimmune disease, cancer.With
Dramatically increasing for the market demand, Antibody preparation production process more and more attention has been paid to.
The downstream preparation process of antibody drug mainly includes two stages of antibody capture and polishing purification.Fermented liquid supernatant liquid
After affinity chromatography, product purity generally can reach 90% monomer purity.But acceptable antibody purity requirement is high, antibody warp
After capture, product composition is still more complex, containing plurality of impurities such as DNA, aggressiveness and host cell proteins, need to generally hand over again through anion
It changes, cation-exchange chromatography and hydrophobic interaction chromatography further clean.But the polishing purification stage needs various column chromatography connection
With encountering the problems such as such as purification process is complicated, step is more or at high cost.Therefore, it is necessary to study the new of antibody polishing purification
Method.
In view of this, the present invention is specifically proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of antibody purification process based on mixed fillers, this method is by by anion
Displacement chromatography medium and hydrophobic chromatoghaphy medium are used in mixed way to obtain mixed fillers, carry out essence to antibody using mixed fillers one-step method
Pure processing, makes full use of that mixed fillers adsorption capacity is big, good selective, and separative efficiency is high, simplifies separating step, reduces
Separation costs.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
Antibody purification process based on mixed fillers, includes the following steps:
Crude product containing target antibody is purified using mixed fillers chromatography column, mixed fillers include anion-exchange chromatography
Medium and hydrophobic chromatoghaphy medium.
Antibody purification process of the invention, by the way that anion-exchange chromatography medium and hydrophobic chromatoghaphy medium to be used in mixed way
To mixed fillers, consummate processing is carried out to antibody using mixed fillers one-step method, makes full use of that mixed fillers adsorption capacity is big, choosing
The advantages that selecting property is good, separative efficiency is high, simplifies separating step, reduces separation costs.
Using mixed fillers, a variety of interactions, more single anion exchange effect or hydrophobic can be generated with albumen
Effect, the target antibody rate of recovery is high, can effectively remove the impurity such as high polymer, solves that cannot be considered in terms of target in the prior art anti-
The problem of body purity and the rate of recovery.
To sum up, compared with prior art, antibody purification process of the invention, operational sequence is simple, passes through one-step method
It realizes to the consummate of target antibody, and adsorption capacity is big, treating capacity is high, antibody purity is high, and the rate of recovery is high.
Preferably, the volume ratio of anion-exchange chromatography medium and hydrophobic chromatoghaphy medium is 5 ﹕, 1~1 ﹕ 5,1~1 ﹕ of preferably 3 ﹕
3。
Preferably, anion-exchange chromatography medium includes appointing in POROS 50HQ, Capto Q and Capto Q ImpRes
It is one or more, preferably POROS 50HQ.
Preferably, hydrophobic chromatoghaphy medium includes in Capto Butyl ImpRes, Capto Octyl and POROS Ethyl
Any one or more, preferably Capto Butyl ImpRes.
POROS 50HQ medium is that a kind of strong anion exchange of Thermo Fisher Scientific company exploitation is situated between
Matter, using styrene diethylene benzene copoly mer microballoon as matrix, Quaternary Polyethyleneimine is aglucon.Capto Butyl
ImpRes medium is a kind of hydrophobic chromatoghaphy medium of GE company exploitation, and using agarose microbeads as matrix, butyl is aglucon.Using two
Kind medium compounding, can give full play to anion exchange effect and hydrophobic effect, further increase suction of the mixed fillers to impurity
Attached effect improves removal of impurity, while improving the elution efficiency of target antibody, guarantees the rate of recovery.
Preferably, the equilibration buffer used is purified as Tris- acetate buffer solution.It is furthermore preferred that the pH of equilibration buffer
For 7.1-7.9, preferably 7.3-7.6, more preferably 7.5；The concentration of equilibration buffer be 45-55mM, preferably 50-55mM,
More preferably 50mM.
It can be guaranteed using above-mentioned equilibration buffer, the impurity absorptions such as high polymer are in chromatographic column, and target antibody flows through, from
And the purity for flowing through target antibody in liquid is improved, yield is improved, while improving removal of impurity.
Purification process of the invention is primarily adapted for use in the purifying of the purifying of antibody, especially monoclonal antibody, especially Chinese hamster
The purifying of gonad cell expressing recombinant antibody.When target antibody is Chinese hamster ovary cell expression recombinant antibodies, balance is slow
Fliud flushing is the buffer of Tris- acetic acid, pH7.1-7.9, and preferred concentration is 50-55mM Tris- acetate buffer solution.
Preferably, the dress pillar height degree of mixed fillers is 4.8-6.0cm, preferably 5.0-5.4cm, more preferable 5.2cm.
Preferably, the operation linear flow rate of purification process be 80-160cm/h, preferably 100-120cm/h, more preferably
110cm/h。
The dress pillar height degree and flow velocity of filler are the key parameters for influencing purification efficiency and purity, optimized, using the above item
Part purification efficiency increases substantially, and the unit time treating capacity of unit packing volume significantly improves, and combines and guarantees high mesh
Labeling antibody purity.
Preferably, the preparation method of the crude product containing target antibody includes: the cell fermentation liquid containing target antibody, through deep layer mistake
Filter, protein A affinity chromatography, are virus inactivated, and after adjusting pH to 7.5 ± 0.5, carry out in-depth filtration, obtain containing target antibody
Crude product.The method of protein A affinity chromatography includes, and by albumin A affinity capture, is then washed target antibody with acidic buffer
De-, collection obtains the eluent containing target antibody.Acid solution is added to the eluent containing target antibody, by the pH tune of eluent
To 3.5-3.7, after 25 DEG C of holding about 2h, in alkaline solution and eluent pH to 7.0-7.5 or so, further deep layer mistake
Filter, obtains clear crude product solution containing target antibody.
Preferably, when preparing crude product solution containing target antibody, acetic acid-that the acidic buffer used is 3.7 ± 0.3 for pH
Sodium acetate buffer, preferably 3.7.
Preferably, the acid solution that adjusting eluent pH to 3.5-3.7 is used neutralizes eluent pH extremely for acetum
The alkaline solution that 7.0-7.5 is used is Tris solution.
The in-depth filtration that the present invention uses, preferably two-stage in-depth filtration, including primary in-depth filtration and second level deep layer mistake
Filter.Two-stage in-depth filtration further preferably is carried out using two concatenated filters, the solution containing target antibody is by primary deep
After layer filter filtering, it is filtered using second level deep bed filter.Millipore can be selected in primary deep bed filter
Millipore A1HC can be selected in D0HC, second level deep bed filter.
Compared with prior art, the invention has the benefit that
(1) antibody purification process of the invention is made by mixing anion-exchange chromatography medium with hydrophobic chromatoghaphy medium
With mixed fillers are obtained, consummate processing is carried out to antibody using mixed fillers one-step method, makes full use of mixed fillers adsorption capacity
Greatly, good selective, separative efficiency is high, simplifies separating step, reduces separation costs；
(2) present invention uses mixed fillers, and a variety of interactions can be generated with albumen, and more single anion exchange is made
With or hydrophobic effect, the target antibody rate of recovery is high, can effectively remove the impurity such as high polymer, solving in the prior art can not be simultaneous
The problem of caring for target antibody purity and the rate of recovery；
(3) by other technological parameters in optimized purification method, the removal rate to impurity is further improved, improves mesh
Labeling antibody purity, and improve the rate of recovery.
Specific embodiment
Technical solution of the present invention is clearly and completely described below in conjunction with specific embodiment, but ability
Field technique personnel will be understood that following described embodiments are some of the embodiments of the present invention, instead of all the embodiments,
It is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.Based on the embodiments of the present invention, the common skill in this field
Art personnel every other embodiment obtained without making creative work belongs to the model that the present invention protects
It encloses.The person that is not specified actual conditions in embodiment, carries out according to conventional conditions or manufacturer's recommended conditions.Agents useful for same or instrument
Production firm person is not specified, is the conventional products that can be obtained by commercially available purchase.
Embodiment 1
The antibody purification process based on mixed fillers of the present embodiment, includes the following steps:
Sample source is Chinese hamster ovary cell expression recombinant antibodies.
The cell fermentation liquid containing monoclonal antibody is taken, Millipore D0HC and the Millipore A1HC two being concatenated
Grade deep bed filter carries out in-depth filtration removal cell fragment and partial impurities, is then captured by affine resin, using acetic acid-
Sodium acetate, pH3.7 acidic buffer solution target antibody is eluted, collect eluent；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
It pre-installs chromatographic column (Millipore Vantage), internal diameter 1.1cm, the volume ratio for filling 5.2mL is the POROS of 3 ﹕ 1
50HQ and Capto Butyl ImpRes mixed fillers medium, operation linear flow rate are 110cm/h.Use equilibrating buffer
(50mM Tris-HAc, pH 7.5) balances chromatographic column, then loading, and sample source is the crude product containing target antibody that upper step obtains
Solution starts to penetrate to sample, and collection flows through liquid, obtains monoclonal antibody sterling, and SEC purity assay is 99.6%, and yield is
95.7%, high polymer content has dropped 50.8%.
Embodiment 2
The antibody purification process based on mixed fillers of the present embodiment, includes the following steps:
Sample source is Chinese hamster ovary cell expression recombinant antibodies.
The cell fermentation liquid containing monoclonal antibody is taken, Millipore D0HC and the Millipore A1HC two being concatenated
Grade deep bed filter carries out in-depth filtration removal cell fragment and partial impurities, is then captured by affine resin, using acetic acid-
Sodium acetate, pH3.7 acidic buffer solution target antibody is eluted, collect eluent；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
It pre-installs chromatographic column (Millipore Vantage), internal diameter 1.1cm, the volume ratio for filling 5.2mL is the POROS of 1 ﹕ 1
50HQ and Capto Butyl ImpRes mixed fillers medium, operation linear flow rate are 110cm/h.Use equilibrating buffer
(50mM Tris-HAc, pH 7.5) balances chromatographic column, then loading, and sample source is the crude product containing target antibody that upper step obtains
Solution starts to penetrate to sample, and collection flows through liquid, obtains monoclonal antibody sterling, and SEC purity assay is 99.0%, and yield is
95.1%, high polymer content has dropped 55.5%.
Embodiment 3
The antibody purification process based on mixed fillers of the present embodiment, includes the following steps:
Sample source is Chinese hamster ovary cell expression recombinant antibodies.
The cell fermentation liquid containing monoclonal antibody is taken, Millipore D0HC and the Millipore A1HC two being concatenated
Grade deep bed filter carries out in-depth filtration removal cell fragment and partial impurities, is then captured by affine resin, using acetic acid-
Sodium acetate, pH3.7 acidic buffer solution target antibody is eluted, collect eluent；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
It pre-installs chromatographic column (Millipore Vantage), internal diameter 1.1cm, the volume ratio for filling 5.2mL is the POROS of 1 ﹕ 3
50HQ and Capto Butyl ImpRes mixed fillers medium, operation linear flow rate are 110cm/h.Use equilibrating buffer
(50mM Tris-HAc, pH 7.5) balances chromatographic column, then loading, and sample source is the crude product containing target antibody that upper step obtains
Solution starts to penetrate to sample, and collection flows through liquid, obtains monoclonal antibody sterling, and SEC purity assay is 99.7%, and yield is
93.8%, high polymer content has dropped 64.3%.
Embodiment 4
The antibody purification process based on mixed fillers of the present embodiment, includes the following steps:
Sample source is Chinese hamster ovary cell expression recombinant antibodies.
The cell fermentation liquid containing monoclonal antibody is taken, Millipore D0HC and the Millipore A1HC two being concatenated
Grade deep bed filter carries out in-depth filtration removal cell fragment and partial impurities, is then captured by affine resin, using acetic acid-
Sodium acetate, pH3.7 acidic buffer solution target antibody is eluted, collect eluent；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
It pre-installs chromatographic column (Millipore Vantage), internal diameter 1.1cm, the volume ratio for filling 5.2mL is the POROS of 5 ﹕ 1
50HQ and Capto Butyl ImpRes mixed fillers medium, operation linear flow rate are 110cm/h.Use equilibrating buffer
(50mM Tris-HAc, pH 7.5) balances chromatographic column, then loading, and sample source is the crude product containing target antibody that upper step obtains
Solution starts to penetrate to sample, and collection flows through liquid, obtains monoclonal antibody sterling, and SEC purity assay is 98.8%, and yield is
96.8%, high polymer content has dropped 30.6%.
Embodiment 5
The antibody purification process based on mixed fillers of the present embodiment, includes the following steps:
Sample source is Chinese hamster ovary cell expression recombinant antibodies.
The cell fermentation liquid containing monoclonal antibody is taken, Millipore D0HC and the Millipore A1HC two being concatenated
Grade deep bed filter carries out in-depth filtration removal cell fragment and partial impurities, is then captured by affine resin, using acetic acid-
Sodium acetate, pH3.7 acidic buffer solution target antibody is eluted, collect eluent；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
It pre-installs chromatographic column (Millipore Vantage), internal diameter 1.1cm, the volume ratio for filling 5.2mL is the POROS of 1 ﹕ 5
50HQ and Capto Butyl ImpRes mixed fillers medium, operation linear flow rate are 110cm/h.Use equilibrating buffer
(50mM Tris-HAc, pH 7.5) balances chromatographic column, then loading, and sample source is the crude product containing target antibody that upper step obtains
Solution starts to penetrate to sample, and collection flows through liquid, obtains monoclonal antibody sterling, and SEC purity assay is 99.5%, and yield is
90.2%, high polymer content has dropped 71.6%.
Comparative example 1
Sample source is Chinese hamster ovary cell expression recombinant antibodies.The cell fermentation liquid containing monoclonal antibody is taken,
Millipore D0HC and Millipore the A1HC two-stage deep bed filter being concatenated carries out in-depth filtration removal cell fragment
And partial impurities, then by affine resin capture, using Acetic acid-sodium acetate, pH3.7 acidic buffer solution by target antibody
Eluent is collected in elution；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
It pre-installs chromatographic column (Millipore Vantage), internal diameter 1.1cm fills the POROS50HQ medium of 5.2mL, operates
Linear flow rate is 110cm/h.Chromatographic column is balanced with equilibrating buffer (50mM Tris-HAc, pH 7.5), then loading, sample
Product source is the crude product solution containing target antibody that upper step obtains, and starts to penetrate to sample, and collection flows through liquid, obtains monoclonal antibody
Sterling, SEC purity assay are 98.2%, and yield 98.7%, high polymer content has dropped 13.7%.
Comparative example 2
Sample source is Chinese hamster ovary cell expression recombinant antibodies.The cell fermentation liquid containing monoclonal antibody is taken,
Millipore D0HC and Millipore the A1HC two-stage deep bed filter being concatenated carries out in-depth filtration removal cell fragment
And partial impurities, then by affine resin capture, using Acetic acid-sodium acetate, pH3.7 acidic buffer solution by target antibody
Eluent is collected in elution；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
It pre-installs chromatographic column (Millipore Vantage), internal diameter 1.1cm fills the Capto Butyl ImpRes of 5.2mL
Medium, operation linear flow rate are 110cm/h.Chromatographic column is balanced with equilibrating buffer (50mM Tris-HAc, pH 7.5), so
Loading afterwards, sample source are the crude product solution containing target antibody that upper step obtains, and start to penetrate to sample, and collection flows through liquid, are obtained
Monoclonal antibody sterling, SEC purity assay are 99.4%, and yield 87.5%, high polymer content has dropped 75.7%.
Comparative example 3
Sample source is Chinese hamster ovary cell expression recombinant antibodies.The cell fermentation liquid containing monoclonal antibody is taken,
Millipore D0HC and Millipore the A1HC two-stage deep bed filter being concatenated carries out in-depth filtration removal cell fragment
And partial impurities, then by affine resin capture, using Acetic acid-sodium acetate, pH3.7 acidic buffer solution by target antibody
Eluent is collected in elution；
Acetum is added into eluent, adjusts the pH to 3.7 of eluent, after keeping 2h under the conditions of 25 DEG C, uses
It is obtained clear containing mesh with eluent pH to 7.5 further by two-stage deep bed filter progress in-depth filtration in Tris solution
Labeling antibody crude product solution.
Pre-install chromatographic column (Millipore Vantage), internal diameter 1.1cm, respectively fill 5.2mL POROS 50HQ and
Capto Butyl ImpRes medium, is used in series, and operation linear flow rate is 110cm/h.With equilibrating buffer (50mM
Tris-HAc, pH 7.5) balance chromatographic column, then loading, sample source are the crude product solution containing target antibody that upper step obtains, to
Sample starts to penetrate, and collection flows through liquid, obtains monoclonal antibody sterling, and SEC purity assay is 99.6%, yield 85.3%,
High polymer content has dropped 78.9%.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: its according to
So be possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features into
Row equivalent replacement；And these are modified or replaceed, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. the antibody purification process based on mixed fillers, which comprises the steps of:
Crude product containing target antibody is purified using mixed fillers chromatography column, the mixed fillers include anion-exchange chromatography
Medium and hydrophobic chromatoghaphy medium.
2. the antibody purification process according to claim 1 based on mixed fillers, which is characterized in that the anion exchange
The volume ratio of chromatography media and the hydrophobic chromatoghaphy medium is 5 ﹕, 1~1 ﹕ 5；
Preferably, the volume ratio of the anion-exchange chromatography medium and the hydrophobic chromatoghaphy medium is 3 ﹕, 1~1 ﹕ 3.
3. the antibody purification process according to claim 1 based on mixed fillers, which is characterized in that the anion exchange
Chromatography media includes POROS 50HQ, Capto Q and Capto Q ImpRes any one or more of；
Preferably, the anion-exchange chromatography medium is POROS 50HQ.
4. the antibody purification process according to claim 1 based on mixed fillers, which is characterized in that the hydrophobic chromatography is situated between
Matter includes Capto Butyl ImpRes, Capto Octyl and Poros Ethyl any one or more of；
Preferably, the hydrophobic chromatoghaphy medium is Capto Butyl ImpRes.
5. the antibody purification process according to claim 1 based on mixed fillers, which is characterized in that in the purifying, adopt
Equilibration buffer is Tris- acetate buffer solution；
Preferably, the pH of the equilibration buffer is 7.1-7.9；
It is furthermore preferred that the pH of the equilibration buffer is 7.3-7.6.
6. the antibody purification process according to claim 5 based on mixed fillers, which is characterized in that the equilibration buffer
For 45-55mM Tris- acetate buffer solution；
It is furthermore preferred that the equilibration buffer is 50-55mM Tris- acetate buffer solution.
7. the antibody purification process according to claim 1 based on mixed fillers, which is characterized in that the mixed fillers
Dress pillar height degree is 4.8-6.0cm；
Preferably, the filling height of the mixed fillers is 5.0-5.4cm.
8. the antibody purification process according to claim 1 based on mixed fillers, which is characterized in that the operation of the purifying
Linear flow rate is 80-160cm/h；
Preferably, the operation linear flow rate of the purifying is 100-120cm/h.
9. the antibody purification process according to claim 1 based on mixed fillers, which is characterized in that described to contain target antibody
The preparation method of crude product includes: the cell fermentation liquid containing target antibody, through in-depth filtration, protein A affinity chromatography, carries out virus and goes out
It is living, after adjusting pH to 7.5 ± 0.5, in-depth filtration is carried out, obtains the crude product containing target antibody.
10. the antibody purification process according to claim 9 based on mixed fillers, which is characterized in that the in-depth filtration
For two-stage in-depth filtration；
Preferably, the in-depth filtration includes carrying out two-stage deep layer by Millipore D0HC and Millipore A1HC respectively
Filtering.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111499682A (en) * | 2020-04-23 | 2020-08-07 | 杭州奕安济世生物药业有限公司 | Clarification treatment method of cell fermentation liquor and separation and purification method of antibody |
| CN113929733A (en) * | 2021-10-25 | 2022-01-14 | 江苏帆博生物制品有限公司 | Monoclonal antibody purification method of mixed ion exchange filler |
| CN114113414A (en) * | 2021-12-23 | 2022-03-01 | 宁波市疾病预防控制中心 | Polymer composite microsphere for extracting and purifying triphenyl metabolite in urine, preparation method, kit and extraction method |
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