title: Biochemical and biophysical characterization of humanized IgG1 produced in Pichia pastoris.
abstract: The first full length IgG produced in Pichia pastoris was reported in late 1980. However, use of a wild-type Pichia expression system to produce IgGs with human-like N-linked glycans was not possible until recently. Advances in glycoengineering have enabled organisms such as Pichia to mimic human N-glycan biosynthesis and produce IgGs with human glycans on an industrial scale. Since there are only a few reports of the analytical characterization of Pichia-produced IgG, we summarize the results known in this field, and provide additional characterization data generated in our laboratories. The data suggest that Pichia-produced IgG has the same stability as that produced in Chinese hamster ovary (CHO) cells. It has similar aggregation profiles, charge variant distribution and oxidation levels as those for a CHO IgG. It contains human N-linked glycans and O-linked single mannose. Because of the comparable biophysical and biochemical characteristics, glycoengineered Pichia pastoris is an attractive expression system for therapeutic IgG productions.
authors: Department of Bioprocess Analytical and Formulation Sciences, Merck Research Laboratories, West Point, PA, USA.
authors: Sha
authors: S
authors: Ha
authors: Yang
authors: Y
authors: Wang
authors: Richard R
authors: RR
authors: Rustandi
mesh_terms: D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity / Q000502:physiology; D016466:CHO Cells; D006224:Cricetinae; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D007074:Immunoglobulin G; D011994:Recombinant Proteins