title: Glycoengineered Pichia produced anti-HER2 is comparable to trastuzumab in preclinical study.
abstract: Mammalian cell culture systems are used predominantly for the production of therapeutic monoclonal antibody (mAb) products. A number of alternative platforms, such as Pichia engineered with a humanized N-linked glycosylation pathway, have recently been developed for the production of mAbs. The glycosylation profiles of mAbs produced in glycoengineered Pichia are similar to those of mAbs produced in mammalian systems. This report presents for the first time the comprehensive characterization of an anti-human epidermal growth factor receptor 2 (HER2) mAb produced in a glycoengineered Pichia, and a study comparing the anti-HER2 from Pichia, which had an amino acid sequence identical to trastuzumab, with trastuzumab. The comparative study covered a full spectrum of preclinical evaluation, including bioanalytical characterization, in vitro biological functions, in vivo anti-tumor efficacy and pharmacokinetics in both mice and non-human primates. Cell signaling and proliferation assays showed that anti-HER2 from Pichia had antagonist activities comparable to trastuzumab. However, Pichia-produced material showed a 5-fold increase in binding affinity to FcγIIIA and significantly enhanced antibody dependant cell-mediated cytotoxicity (ADCC) activity, presumably due to the lack of fucose on N-glycans. In a breast cancer xenograft mouse model, anti-HER2 was comparable to trastuzumab in tumor growth inhibition. Furthermore, comparable pharmacokinetic profiles were observed for anti-HER2 and trastuzumab in both mice and cynomolgus monkeys. We conclude that glycoengineered Pichia provides an alternative production platform for therapeutic mAbs and may be of particular interest for production of antibodies for which ADCC is part of the clinical mechanism of action.
authors.affiliation: Department of Biologics Research; Merck Research Laboratory, West Point, PA, USA.
authors.forename: Ningyan
authors.initials: N
authors.lastname: Zhang
authors.forename: Liming
authors.initials: L
authors.lastname: Liu
authors.forename: Calin Dan
authors.initials: CD
authors.lastname: Dumitru
authors.forename: Nga Rewa Houston
authors.initials: NR
authors.lastname: Cummings
authors.forename: Michael
authors.initials: M
authors.lastname: Cukan
authors.forename: Youwei
authors.initials: Y
authors.lastname: Jiang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.forename: Teresa
authors.initials: T
authors.lastname: Mitchell
authors.forename: Muralidhar R
authors.initials: MR
authors.lastname: Mallem
authors.forename: Yangsi
authors.initials: Y
authors.lastname: Ou
authors.forename: Rohan N
authors.initials: RN
authors.lastname: Patel
authors.forename: Kim
authors.initials: K
authors.lastname: Vo
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Irina
authors.initials: I
authors.lastname: Burnina
authors.forename: Byung-Kwon
authors.initials: BK
authors.lastname: Choi
authors.forename: Hans E
authors.initials: HE
authors.lastname: Huber
authors.forename: Terrance A
authors.initials: TA
authors.lastname: Stadheim
authors.forename: Dongxing
authors.initials: D
authors.lastname: Zha
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D061067:Antibodies, Monoclonal, Humanized / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000920:Antibody-Dependent Cell Cytotoxicity / Q000187:drug effects / Q000276:immunology; D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000494:pharmacology; D019540:Area Under Curve; D001667:Binding, Competitive / Q000276:immunology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D005643:Fucose / Q000378:metabolism; D005818:Genetic Engineering; D006801:Humans; D008252:Macaca fascicularis; D051379:Mice; D008810:Mice, Inbred C57BL; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D011485:Protein Binding / Q000276:immunology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D017452:Receptors, IgG / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology* / Q000493:pharmacokinetics / Q000494:pharmacology; D000068878:Trastuzumab; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; D005643:Fucose; D011134:Polysaccharides; D018719:Erb-b2 Receptor Tyrosine Kinases; D017452:Receptors, IgG; D011994:Recombinant Proteins; D000068878:Trastuzumab; C496124:FCGR3A protein, human