title: Evaluation of heavy chain C-terminal deletions on productivity and product quality of monoclonal antibodies in Chinese hamster ovary (CHO) cells.
abstract: Monoclonal antibodies (mAbs) have been well established as potent therapeutic agents and are used to treat many different diseases. During cell culture production, antibody charge variants can be generated by cleavage of heavy chain (HC) C-terminal lysine and proline amidation. Differences in levels of charge variants during manufacturing process changes make it challenging to demonstrate process comparability. In order to reduce heterogeneity and achieve consistent product quality, we generated and expressed antibodies with deletion of either HC C-terminal lysine (-K) or lysine and glycine (-GK). Interestingly, clones that express antibodies lacking HC C-terminal lysine (-K) had considerably lower specific productivities compared to clones that expressed either wild type antibodies (WT) or antibodies lacking HC glycine and lysine (-GK). While no measurable differences in antibody HC and LC mRNA levels, glycosylation and secretion were observed, our analysis suggests that the lower specific productivity of clones expressing antibody lacking HC C-terminal lysine was due to slower antibody HC synthesis and faster antibody degradation. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:786-794, 2017.
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Zhilan
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Danming
authors.identifier: http://orcid.org/0000-0002-7199-743X
authors.initials: D
authors.lastname: Tang
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Shahram
authors.initials: S
authors.lastname: Misaghi
authors.affiliation: Biological Technologies, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Guoying
authors.initials: G
authors.lastname: Jiang
authors.affiliation: Protein Analytical Chemistry, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Christopher
authors.initials: C
authors.lastname: Yu
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Mandy
authors.initials: M
authors.lastname: Yim
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: David
authors.initials: D
authors.lastname: Shaw
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Brad
authors.initials: B
authors.lastname: Snedecor
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Laird
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc, 1 DNA Way, South San Francisco, CA, 94080.
authors.forename: Amy
authors.initials: A
authors.lastname: Shen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005998:Glycine / Q000378:metabolism; D006031:Glycosylation; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D011392:Proline / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D012333:RNA, Messenger; D011392:Proline; D008239:Lysine; D005998:Glycine
keywords: C-terminal lysine; CHO cell; charge variants; productivity; proline amidation; therapeutic monoclonal antibody