title: A high-throughput purification of monoclonal antibodies from glycoengineered Pichia pastoris.
abstract: 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.
authors.affiliation: GlycoFi, Inc., 21 Lafayette Street, Suite 200, Lebanon, NH 03766, USA.
authors.forename: Youwei
authors.initials: Y
authors.lastname: Jiang
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.forename: Michelle
authors.initials: M
authors.lastname: Button
authors.forename: Michael
authors.initials: M
authors.lastname: Cukan
authors.forename: Renee
authors.initials: R
authors.lastname: Moore
authors.forename: Nathan
authors.initials: N
authors.lastname: Sharkey
authors.forename: Huijuan
authors.initials: H
authors.lastname: Li
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods; D005285:Fermentation; D015870:Gene Expression; D006031:Glycosylation; D007074:Immunoglobulin G / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007218:Industrial Microbiology / Q000191:economics / Q000379:methods*; D010843:Pichia / Q000235:genetics*; D015203:Reproducibility of Results; D013205:Staphylococcal Protein A / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A