Evaluation of Heavy-Chain C-Terminal Deletion on Product Quality and Pharmacokinetics of Monoclonal Antibodies.
Due to their potential influence on stability, pharmacokinetics, and product consistency, antibody charge variants have attracted considerable attention in the biotechnology industry. Subtle to significant differences in the level of charge variants and new charge variants under various cell culture conditions are often observed during routine manufacturing or process changes and pose a challenge when demonstrating product comparability. To explore potential solutions to control charge heterogeneity, monoclonal antibodies (mAbs) with native, wild-type C-termini, and mutants with C-terminal deletions of either lysine or lysine and glycine were constructed, expressed, purified, and characterized in vitro and in vivo. Analytical and physiological characterization demonstrated that the mAb mutants had greatly reduced levels of basic variants without decreasing antibody biologic activity, structural stability, pharmacokinetics, or subcutaneous bioavailability in rats. This study provides a possible solution to mitigate mAb heterogeneity in C-terminal processing, improve batch-to-batch consistency, and facilitate the comparability study during process changes.
Biological Technologies, Genentech, 1 DNA Way, South San Francisco, California 94080. Electronic address: jiang.guoying@gene.com.
Guoying
G
Jiang
Protein Analytical Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080.
Christopher
C
Yu
Department of Pharmacokinetic and Pharmacodynamic Sciences, Genentech, 1 DNA Way, South San Francisco, California 94080.
Daniela B
DB
Yadav
Early Stage Cell Culture, Genentech, 1 DNA Way, South San Francisco, California 94080.
Zhilan
Z
Hu
Purification Development, Genentech, 1 DNA Way, South San Francisco, California 94080.
Annamarie
A
Amurao
Purification Development, Genentech, 1 DNA Way, South San Francisco, California 94080.
Eileen
E
Duenas
Purification Development, Genentech, 1 DNA Way, South San Francisco, California 94080.
Marc
M
Wong
Purification Development, Genentech, 1 DNA Way, South San Francisco, California 94080.
Mark
M
Iverson
Late Stage Pharmaceutical and Processing Development, Genentech, 1 DNA Way, South San Francisco, California 94080.
Kai
K
Zheng
Late Stage Pharmaceutical and Processing Development, Genentech, 1 DNA Way, South San Francisco, California 94080.
Xanthe
X
Lam
Protein Analytical Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080.
Jia
J
Chen
BioAnalytical Sciences, Genentech, 1 DNA Way, South San Francisco, California 94080.
Roxanne
R
Vega
BioAnalytical Sciences, Genentech, 1 DNA Way, South San Francisco, California 94080.
Sheila
S
Ulufatu
BioAnalytical Sciences, Genentech, 1 DNA Way, South San Francisco, California 94080.
Cecilia
C
Leddy
BioAnalytical Sciences, Genentech, 1 DNA Way, South San Francisco, California 94080.
Helen
H
Davis
Early Stage Cell Culture, Genentech, 1 DNA Way, South San Francisco, California 94080.
Amy
A
Shen
Biological Technologies, Genentech, 1 DNA Way, South San Francisco, California 94080.
Pin Y
PY
Wong
Protein Analytical Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080.
Reed
R
Harris
Late Stage Pharmaceutical and Processing Development, Genentech, 1 DNA Way, South San Francisco, California 94080.
Y John
YJ
Wang
Department of Pharmacokinetic and Pharmacodynamic Sciences, Genentech, 1 DNA Way, South San Francisco, California 94080.
Dongwei
D
Li
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000493:pharmacokinetics*; D001682:Biological Availability; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004355:Drug Stability; D005998:Glycine / Q000737:chemistry; D007279:Injections, Subcutaneous; D007525:Isoelectric Focusing; D008239:Lysine / Q000737:chemistry; D008297:Male; D009154:Mutation; D051381:Rats; D017207:Rats, Sprague-Dawley
D000911:Antibodies, Monoclonal; D008239:Lysine; D005998:Glycine
C-terminal Lys deletion; Lys and Gly deletion; bioavailability; charge heterogeneity; mAb; pharmacokinetics