A high-throughput purification of monoclonal antibodies from glycoengineered Pichia pastoris

Glycoengineered Pichia pastoris provides a unique platform for screening monoclonal antibody (mAb) leads and high expressing strains. A simple, economic, and high-throughput purification for mAb from P. pastoris fermentation has been developed that can be easily operated in various commercially avai...

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Bibliographic Details
Published in:Protein expression and purification 2010-11, Vol.74 (1), p.9-15
Main Authors: Jiang, Youwei, Li, Fang, Button, Michelle, Cukan, Michael, Moore, Renee, Sharkey, Nathan, Li, Huijuan
Format: Article
Language:English
Subjects:
Antibodies, Monoclonal - genetics
Antibodies, Monoclonal - isolation & purification
Antibodies, Monoclonal - metabolism
Chromatography, High Pressure Liquid - economics
Chromatography, High Pressure Liquid - methods
Fermentation
Gene Expression
Glycosylation
High-throughput
Immunoglobulin G - genetics
Immunoglobulin G - isolation & purification
Immunoglobulin G - metabolism
Industrial Microbiology - economics
Industrial Microbiology - methods
Monoclonal antibody
Pichia - genetics
Pichia pastoris
Purification
Reproducibility of Results
Staphylococcal Protein A - metabolism
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Summary:Glycoengineered Pichia pastoris provides a unique platform for screening monoclonal antibody (mAb) leads and high expressing strains. A simple, economic, and high-throughput purification for mAb from P. pastoris fermentation has been developed that can be easily operated in various commercially available liquid handlers. The method includes the use of STREAMLINE rProtein A in a 96-well platform and demonstrates good linear alignment and reproducibility in a wide concentration range. The antibody titers measured by the method have less than 15% variation in comparison to spiking titers. The mAb titer and quality obtained from this method are comparable to that from conventional column chromatography. The method can process hundreds of expression screening samples in a day, not only to accurately determine titers, but also to generate milligram quantities of mAb for quality assessment, including purity, folding, glycosylation, and antigen binding affinity.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2010.04.016