CN107033236A - A kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth - Google Patents
A kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth Download PDFInfo
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Abstract
The invention discloses a kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth, the protein chromatographic isolation technics belonged in biological chemical field.Comprise the following steps that：1) fermentation liquor pretreatment, takes the yeast fermentation broth containing human serum albumin, and centrifugation removes yeast cells, adds Sodium Caprylate, and heating removes foreign protein and inactivated proteases, centrifuges, take supernatant；2) column chromatography, uses tryptophan for the mixed mode medium of aglucon, fixed bed chromatography supernatant, loading pH value 4.0~4.5, elution pH value 7.0~9.0, collects eluting peak.3) desalination and drying, by collection liquid desalination, freeze-drying obtains the human serum albumin that purity is more than 95%.The present invention characteristic be to develop a kind of new separating technology, the human serum albumin of high-purity can be directly isolated to obtain from yeast fermentation broth.The key of method is to employ the mixed mode medium using tryptophan as aglucon, yeast fermentation broth need not adjust ionic strength, adsorbed under acid condition, the elution of neutral and alkalescent, with elution requirement is gentle, technique is simple, separative efficiency is high, high income the characteristics of.
Description
Technical field
The invention belongs to protein stripping technique field, it is related to a kind of the mixed of separation human serum albumin from yeast fermentation broth
Syntype chromatography method.
Background technology
Human serum albumin is content most abundant protein in human plasma, accounts for plasma protein 60%, major physiological work(
Can be maintain plasma osmolarity, with reference to and transport nutriment.It is clinically used for treatment haemorrhagic shock, traumatic shock, urgency
Property hypovolemia and hypoalbuminemia etc..Adding ingredient, pharmaceutical adjuvant, excipient of cell culture etc. are alternatively arranged as, is had
Have a wide range of applications.
The human serum albumin product of the Clinical practice mainly extraction purification from the blood plasma of people source, preparation method includes cold ethanol
Precipitation, ammonium sulfate precipitation, rivanol precipitation, caprylate precipitation, chromatography etc., raw material is nervous, and separation is complicated, and can not exclude virus
Or the influence of other potential virulence factors.The yeast cells of expression recombinant human serum albumin is constructed using modern biotechnology, can
Viral infection is avoided, the problems such as blood source is in short supply is solved, has a good application prospect.But, medicinal human serum albumin
Purity requirement is high, and yeast fermentation broth complicated components, contains protease, foreign protein, nucleic acid, aliphatic acid, pigment, polysaccharide and pyrogen
The plurality of impurities such as material, it is necessary to could be removed using more chromatographic enrichment method.
Yeast fermentation broth general pH about 6.0, conductance is 15~25mS/cm.Patent CN1127299 and US5521287 are utilized
Heating, dilution, acid adjustment, cation-exchange chromatography, hydrophobic interaction chromatography, metal chelate chromatography, anion-exchange chromatography, line borate
The steps such as precipitation, obtain the higher human serum albumin of purity.Patent US5962649 substitutes sun with cation exchange Expanded Bed Adsorption
Ion-exchange chromatography, shortens technique.But, both the above method captures human serum albumin using cation-exchange chromatography,
Separation selectivity is limited, while feed liquid needs dilution to handle so that target concentration declines, and adsorption capacity reduction, treating capacity increases
Greatly, process time extends.Patent CN1854155 discloses a kind of Capto MMC media and captures the white egg of human blood in acid condition
In vain, the removal of impurity is gone in high salt elution, neutral and add and afford human serum albumin, the albumin yield that this method is obtained under the conditions of salt
For 65.7%.Keep patent CN1854155 loading and elution processes constant in patent CN 101768206A, remove middle drench
Step is washed, it is 92% to obtain human serum albumin purity, and yield is 71.3%.Patent CN101260145, which is also announced, uses Capto
MMC separates the chromatographic purifying technique of recombinant human serum albumin and its fusion protein, and yield and purity are more than 80%.Above-mentioned base
There are Similar Problems in Capto MMC patent, purity and yield are generally relatively low, some processes need high salt elution to go the removal of impurity
Absorption, process is complicated.The characteristics of for yeast fermentation broth, the capture rate of raising human serum albumin, a kind of high selectivity of exploitation,
With salt-tolerant trait, high income, the simple novel method for separating of process, it will help simplify the separating technology of human serum albumin, carry
High efficiency, it is cost-effective.
Mixed-Modechromatography is a kind of new bioseparation technology, and aglucon has a variety of functional groups concurrently, can be with target
Albumen produces a variety of interactions, mainly hydrophobic and electrostatic interaction.The ligand density of mixed mode medium is generally higher,
Adsorption capacity is big, with salt tolerant characterization of adsorption, and elution requirement is gentle, is particularly suitable for isolating and purifying on a large scale, in antibody
Deng being applied in the isolating and purifying of albumen.MX-Trp-650M media are the hybrid guided modes of TOSOH BIOSCIENCE companies exploitation
Formula medium, using polymethacrylates microballoon as matrix, the amino of tryptophan is coupled in matrix by amido link is used as function
Aglucon, ligand structure is shown in Fig. 1, is mainly used in the polishing purification of antibody, such as patent US0264685A1.Consult domestic and international patent and
Document, does not find that MX-Trp-650M and its similar mediums are used to separate human serum albumin from yeast fermentation broth
The present invention utilizes the advantage of Mixed-Modechromatography, and the particularity that human serum albumin and small molecule are combined, and selects
Using tryptophan as the mixed mode medium of main functional ligand, human serum albumin, exploitation one are directly captured from yeast fermentation broth
Plant the new method that Mixed-Modechromatography separates human serum albumin.
The content of the invention
It is an object of the invention to provide a kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth,
Specifically include following steps：
1) yeast fermentation broth containing human serum albumin is taken, yeast cells is removed by centrifuging, pH to 6.0 is adjusted, added
Sodium Caprylate centrifuges 10~20min with 8000~10000rpm rotating speeds with centrifuge, taken to 5~20mM, 68 DEG C of heating 30min
Clear liquid, obtains human serum albumin crude product solution；
2) pH to 4.0~4.5 of human serum albumin crude product solution is adjusted, after 0.45 μm of membrane filtration, filling is loaded to
In the chromatographic column for having the mixed mode medium using tryptophan as aglucon, Equilibration buffer wash, elution buffer elution, collection is washed
The de- corresponding elution buffer in peak, obtains human serum albumin solution；
3) by human serum albumin solution desalination, freeze-drying obtains the human serum albumin that purity is more than 95%；
The described yeast fermentation broth containing human serum albumin is the zymotic fluid of Pichia anomala expression recombination human blood albumin,
Or saccharomyces cerevisiae expresses the zymotic fluid of recombinant human serum albumin；
Described mixed mode medium is MX-Trp-650M, or is used as aglucon using amido link coupling tryptophan amino
Chromatography media；
Described level pad is acetic acid-sodium acetate buffer solution, and pH value is 4.0~4.5；
Described elution buffer is sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, Tris- hydrochloride buffers or sweet ammonia
Acid-sodium hydrate buffer solution, pH value is 7.0~9.0, addition 0-0.6M NaCl；
Described step 1) in regulation yeast fermentation broth pH value and step 3) human serum albumin crude product solution pH regulation institute
It is acetic acid or citric acid solution with acid solution, aqueous slkali used is sodium hydroxide solution.
The characteristics of present invention is directed to yeast fermentation broth complicated components, conductance is higher, is directly handled using mixed mode medium
Pretreated yeast fermentation broth, make full use of that the adsorption capacity of Mixed-Modechromatography is big, selectivity is good, with salt-tolerant trait,
The advantage such as elution requirement is gentle, improves separative efficiency, simplifies separating step, obtain one and people is directly separated from yeast fermentation broth
The new method of blood albumin.The advantage of the invention is that：1) the mixed mode medium using tryptophan as main functional ligand is used,
Using the specific binding of tryptophan and human serum albumin, the adsorptive selectivity of human serum albumin is improved, single step chromatography is pure
Degree up to more than 95%, pigment removal is up to more than 90%；2) mixed mode medium has good salt tolerant characterization of adsorption, feed liquid
Ionic strength need not be adjusted, direct loading simplifies pre-treatment step, improves separative efficiency；
3) adsorption capacity of mixed mode medium is big, and applied sample amount reaches more than 40mg/ml media, and process treating capacity is big；4)
Elution requirement gently, under pH neutrality or weak basic condition, utilizes electrostatic repulsion to realize effective elution, time of human serum albumin
High income, reaches more than 92%；5) technique is simple, it is easy to amplify.
Brief description of the drawings
Fig. 1 is the ligand structure schematic diagram of mixed mode medium.
Fig. 2 is the chromatography spectrogram that Mixed-Modechromatography of the present invention separates human serum albumin.
Fig. 3 is the feed liquid and elution fraction efficient liquid phase chromatographic analysis figure of Mixed-Modechromatography separation of the present invention.
Embodiment
The present invention provides a kind of method that human serum albumin is separated from yeast fermentation broth.Take the ferment containing human serum albumin
Female zymotic fluid, after centrifugal clarification, adds Sodium Caprylate, heating removes foreign protein and inactivated proteases, centrifuges, take
Clear liquid；Supernatant is separated by Mixed-Modechromatography post, human serum albumin is obtained.The human blood that the inventive method is obtained is white
Purity of protein is more than 95%.
The method that human serum albumin is separated from yeast fermentation broth comprises the following steps：
1) yeast fermentation broth containing human serum albumin is taken, yeast cells is removed by centrifuging, pH to 6.0 is adjusted, added
Sodium Caprylate centrifuges 10~20min with 8000~10000rpm rotating speeds with centrifuge, taken to 5~20mM, 68 DEG C of heating 30min
Clear liquid, obtains human serum albumin crude product solution；
2) pH to 4.0~4.5 of human serum albumin crude product solution is adjusted, after 0.45 μm of membrane filtration, filling is loaded to
In the chromatographic column for having the mixed mode medium using tryptophan as aglucon, Equilibration buffer wash, elution buffer elution, collection is washed
The de- corresponding elution buffer in peak, obtains human serum albumin solution；
3) by human serum albumin solution desalination, freeze-drying obtains the human serum albumin that purity is more than 95%；
The described yeast fermentation broth containing human serum albumin is the zymotic fluid of Pichia anomala expression recombination human blood albumin,
Or saccharomyces cerevisiae expresses the zymotic fluid of recombinant human serum albumin；
Described mixed mode medium is MX-Trp-650M, or is used as aglucon using amido link coupling tryptophan amino
Chromatography media；
Described level pad is acetic acid-sodium acetate buffer solution, and pH value is 4.0~4.5；
Described elution buffer is sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, Tris- hydrochloride buffers or sweet ammonia
Acid-sodium hydrate buffer solution, pH value is 7.0~9.0, addition 0-0.6M NaCl；
Described step 1) in regulation yeast fermentation broth pH value and step 3) human serum albumin crude product solution pH regulation institute
It is acetic acid or citric acid solution with acid solution, aqueous slkali used is sodium hydroxide solution.
Embodiment 1
The Pichia pastoris zymotic fluid containing recombinant human serum albumin, electrical conductivity about 24mS/cm are taken, human serum albumin concentration is
10mg/ml.After centrifugal clarification, adjust pH to 6.0, add Sodium Caprylate to 5mM, 68 DEG C of heating 30min, precipitation foreign protein and
Inactivated proteases, centrifuge 20min with 8000rpm rotating speeds, obtain supernatant.It is 4.0,0.45 μm of filter membrane mistake to adjust supernatant pH value
Filter, takes 10ml as sample introduction sample.Filling 2ml MX-Trp-650M media, level pad in chromatographic column (internal diameter 0.5cm)
For 20mM acetic acid-sodium acetate buffer solutions (pH 4.0), eluent is 20mM sodium dihydrogen phosphates-disodium hydrogen phosphate buffer solution (pH
7.0) elution fraction, is collected, desalination, freeze-drying obtains human serum albumin, and HPLC purity assays are 96.6%, and yield is
96.0%, pigment removal is 90.2%.
Embodiment 2
The Pichia pastoris zymotic fluid containing recombinant human serum albumin, electrical conductivity about 24mS/cm are taken, human serum albumin concentration is
10mg/ml.After centrifugal clarification, pH to 6.0 is adjusted, Sodium Caprylate is added to 20mM, 68 DEG C of heating 30min, precipitates foreign protein
And inactivated proteases, 10min is centrifuged with 10000rpm rotating speeds, supernatant is obtained.It is 4.5,0.45 μm of filter to adjust supernatant pH value
Membrane filtration, takes 10ml as sample introduction sample.Filling 2ml MX-Trp-650M media in chromatographic column (internal diameter 0.5cm), balance is slow
Fliud flushing is 20mM acetic acid-sodium acetate buffer solutions (pH 4.5), and eluent is addition 0.2M NaCl 20mM sodium dihydrogen phosphates-phosphorus
Sour disodium hydrogen buffer solution (pH 7.0), collects elution fraction, and desalination, freeze-drying obtains human serum albumin, HPLC purity assays
For 96.7%, yield is 97.8%, and pigment removal is 91.4%.
Embodiment 3
The Pichia pastoris zymotic fluid containing recombinant human serum albumin, electrical conductivity about 24mS/cm are taken, human serum albumin concentration is
10mg/ml.After centrifugal clarification, pH to 6.0 is adjusted, Sodium Caprylate is added to 15mM, 68 DEG C of heating 30min, precipitates foreign protein
And inactivated proteases, 15min is centrifuged with 9000rpm rotating speeds, supernatant is obtained.It is 4.2,0.45 μm of filter membrane to adjust supernatant pH value
Filtering, takes 10ml as sample introduction sample.Filling 2ml MX-Trp-650M media, equalizing and buffering in chromatographic column (internal diameter 0.5cm)
Liquid is 20mM acetic acid-sodium acetate buffer solution (pH 4.2), and eluent is addition 0.4M NaCl 20mM sodium dihydrogen phosphates-phosphorus
Sour disodium hydrogen buffer solution (pH 7.0), collects elution fraction, and desalination, freeze-drying obtains human serum albumin, chromatography spectrogram
As shown in Fig. 2 HPLC purity assays are 98.5%, yield is 97.4%, and pigment removal is 92.5%, and liquid phase analysis spectrogram is such as
Shown in Fig. 3.
Embodiment 4
The Pichia pastoris zymotic fluid containing recombinant human serum albumin, electrical conductivity about 24mS/cm are taken, human serum albumin concentration is
10mg/ml.After centrifugal clarification, pH to 6.0 is adjusted, Sodium Caprylate is added to 15mM, 68 DEG C of heating 30min, precipitates foreign protein
And inactivated proteases, 15min is centrifuged with 9000rpm rotating speeds, supernatant is obtained.It is 4.0,0.45 μm of filter membrane to adjust supernatant pH value
Filtering, takes 10ml as sample introduction sample.Filling 2ml MX-Trp-650M media, equalizing and buffering in chromatographic column (internal diameter 0.5cm)
Liquid is 20mM acetic acid-sodium acetate buffer solution (pH 4.0), and eluent is 50mM Tris- hydrochloride buffers (pH 8.0), is received
Collect elution fraction, desalination, freeze-drying obtains human serum albumin, and HPLC purity assays are 96.8%, and yield is 96.3%, color
Plain clearance is 91.1%.
Embodiment 5
The Pichia pastoris zymotic fluid containing recombinant human serum albumin, electrical conductivity about 24mS/cm are taken, human serum albumin concentration is
10mg/ml.After centrifugal clarification, pH to 6.0 is adjusted, Sodium Caprylate is added to 20mM, 68 DEG C of heating 30min, precipitates foreign protein
And inactivated proteases, 20min is centrifuged with 8000rpm rotating speeds, supernatant is obtained.It is 4.5,0.45 μm of filter membrane to adjust supernatant pH value
Filtering, takes 10ml as sample introduction sample.Filling 2ml MX-Trp-650M media, equalizing and buffering in chromatographic column (internal diameter 0.5cm)
Liquid is 20mM acetic acid-sodium acetate buffer solution (pH 4.5), and eluent is 50mM Glycine-NaOH sodium buffer solutions (pH
9.0) elution fraction, is collected, desalination, freeze-drying obtains human serum albumin, and HPLC purity assays are 95.2%, and yield is
95.3%, pigment removal is 90.2%.
Embodiment 6
The fermentation by saccharomyces cerevisiae liquid containing recombinant human serum albumin, electrical conductivity about 20mS/cm are taken, human serum albumin concentration is
8mg/ml.After centrifugal clarification, adjust pH to 6.0, add Sodium Caprylate to 10mM, 68 DEG C of heating 30min, precipitation foreign protein and
Inactivated proteases, centrifuge 15min with 9000rpm rotating speeds, obtain supernatant.It is 4.0,0.45 μm of filter membrane mistake to adjust supernatant pH value
Filter, takes 10ml as sample introduction sample.Filling 2ml MX-Trp-650M media, level pad in chromatographic column (internal diameter 0.5cm)
For 20mM acetic acid-sodium acetate buffer solution (pH 4.0), eluent is addition 0.6M NaCl 20mM sodium dihydrogen phosphates-phosphoric acid
Disodium hydrogen buffer solution (pH 7.0), collects elution fraction, and desalination, freeze-drying obtains human serum albumin, HPLC purity assays are
96.8%, yield is 97.5%, and pigment removal is 90.0%.
Embodiment 7
The Pichia pastoris zymotic fluid containing recombinant human serum albumin, electrical conductivity about 24mS/cm are taken, human serum albumin concentration is
10mg/ml.After centrifugal clarification, pH to 6.0 is adjusted, Sodium Caprylate is added to 15mM, 68 DEG C of heating 30min, precipitates foreign protein
And inactivated proteases, 15min is centrifuged with 9000rpm rotating speeds, supernatant is obtained.It is 4.0,0.45 μm of filter membrane to adjust supernatant pH value
Filtering, takes 10ml as sample introduction sample.The amino that filling 2ml is coupled tryptophan with amido link in chromatographic column (internal diameter 0.5cm) is made
For the agarose medium of aglucon, level pad is 20mM acetic acid-sodium acetate buffer solution (pH 4.0), and eluent is addition
0.2M NaCl 20mM sodium dihydrogen phosphates-disodium hydrogen phosphate buffer solution (pH 7.0), collect elution fraction, and desalination, freezing is dry
It is dry, human serum albumin is obtained, HPLC purity assays are 96.7%, and yield is 97.0%, and pigment removal is 92.2%.
Embodiment 8
The Pichia pastoris zymotic fluid containing recombinant human serum albumin, electrical conductivity about 24mS/cm are taken, human serum albumin concentration is
10mg/ml.After centrifugal clarification, pH to 6.0 is adjusted, Sodium Caprylate is added to 15mM, 68 DEG C of heating 30min, precipitates foreign protein
And inactivated proteases, 15min is centrifuged with 9000rpm rotating speeds, supernatant is obtained.It is 4.0,0.45 μm of filter membrane to adjust supernatant pH value
Filtering, takes 10ml as sample introduction sample.The amino that filling 2ml is coupled tryptophan with amido link in chromatographic column (internal diameter 0.5cm) is made
For the cellulose media of aglucon, level pad is 20mM acetic acid-sodium acetate buffer solution (pH 4.0), and eluent is addition
0.2M NaCl 20mM sodium dihydrogen phosphates-disodium hydrogen phosphate buffer solution (pH 7.0), collect elution fraction, and desalination, freezing is dry
It is dry, human serum albumin is obtained, HPLC purity assays are 97.2%, and yield is 96.5%, and pigment removal is 91.1%.
Claims (5)
1. a kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth, it is characterised in that including following step
Suddenly：
1) yeast fermentation broth containing human serum albumin is taken, yeast cells is removed by centrifuging, pH to 6.0 is adjusted, octanoic acid is added
Sodium centrifuges 10~20min with 8000~10000rpm rotating speeds with centrifuge, takes supernatant to 5~20mM, 68 DEG C of heating 30min,
Obtain human serum albumin crude product solution；
2) adjust human serum albumin crude product solution pH to 4.0~4.5, after 0.45 μm of membrane filtration, be loaded to filled with
In chromatographic column of the tryptophan for the mixed mode medium of aglucon, eluting peak is collected in Equilibration buffer wash, elution buffer elution
Corresponding elution buffer, obtains human serum albumin solution；
3) by human serum albumin solution desalination, freeze-drying obtains the human serum albumin that purity is more than 95%；
Described mixed mode medium is MX-Trp-650M, or is coupled tryptophan amino as the chromatography of aglucon using amido link
Medium；
Described level pad pH value is 4.0~4.5；
Described elution buffer pH value is 7.0~9.0.
2. a kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth as claimed in claim 1, its
The zymotic fluid that the described yeast fermentation broth containing human serum albumin is Pichia anomala expression recombination human blood albumin is characterised by,
Or saccharomyces cerevisiae expresses the zymotic fluid of recombinant human serum albumin.
3. a kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth as claimed in claim 1, its
It is acetic acid-sodium acetate buffer solution to be characterised by described level pad.
4. a kind of Mixed-Modechromatography method that human serum albumin is separated from yeast fermentation broth as claimed in claim 1, institute
The elution buffer stated is sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, Tris- hydrochloride buffers or glycine-hydroxide
Sodium buffer solution, addition 0-0.6M NaCl.
5. a kind of Expanded Bed Adsorption separation human serum albumin method based on mixed mode as claimed in claim 1, its feature
Be described step 1) in regulation yeast fermentation broth pH value and step 3) human serum albumin crude product solution pH regulations acid used
Solution is acetic acid or citric acid solution, and aqueous slkali used is sodium hydroxide solution.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114885608A (en) * | 2020-12-08 | 2022-08-09 | 通化安睿特生物制药股份有限公司 | Method for purifying recombinant proteins |
| CN116693659A (en) * | 2023-07-11 | 2023-09-05 | 浙江大学 | Two-step mixed mode chromatography method for separating recombinant human serum albumin |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007079886A1 (en) * | 2005-12-22 | 2007-07-19 | Csl Behring Gmbh | Octanoate-reduced human albumin |
| WO2014145744A1 (en) * | 2013-03-15 | 2014-09-18 | Alder Biopharmaceuticals, Inc. | Antibody purification and purity monitoring |
| CN105063142A (en) * | 2015-08-07 | 2015-11-18 | 山西瑞亚力科技有限公司 | Method used for preparing immunoglobulin G Fc fragments with mixed-mode chromatography filling material |
-
2017
- 2017-05-05 CN CN201710317433.3A patent/CN107033236B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007079886A1 (en) * | 2005-12-22 | 2007-07-19 | Csl Behring Gmbh | Octanoate-reduced human albumin |
| WO2014145744A1 (en) * | 2013-03-15 | 2014-09-18 | Alder Biopharmaceuticals, Inc. | Antibody purification and purity monitoring |
| CN105063142A (en) * | 2015-08-07 | 2015-11-18 | 山西瑞亚力科技有限公司 | Method used for preparing immunoglobulin G Fc fragments with mixed-mode chromatography filling material |
Non-Patent Citations (4)
| Title |
|---|
| QI-CI WU ET AL.: "A mixed-mode resin with tryptamine ligand for human serum albumin separation", 《JOURNAL OF CHROMATOGRAPHY A》 * |
| QICI WU ET AL.: "Preparation and evaluation of mixed-mode resins with tryptophan analogues as functional ligands for human serum albumin separation", 《CHINESE JOURNAL OF CHEMICAL ENGINEERING》 * |
| 吴启赐等: "基于色氨酸类似物的混合模式介质和人血白蛋白吸附性能", 《中国生物化学与分子生物学会2016年全国学术会议》 * |
| 褚文宁等: "基于96-孔板高通量筛选的人血白蛋白层析分离优化", 《中国生物化学与分子生物学会2016年全国学术会议》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114885608A (en) * | 2020-12-08 | 2022-08-09 | 通化安睿特生物制药股份有限公司 | Method for purifying recombinant proteins |
| CN116693659A (en) * | 2023-07-11 | 2023-09-05 | 浙江大学 | Two-step mixed mode chromatography method for separating recombinant human serum albumin |
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