Characterization of the basic charge variants of a human IgG1 Effect of copper concentration in cell culture media

被引：87
|
作者：
Kaschak, Timothy [1 ]
Boyd, Daniel [1 ]
Lu, Franklin [1 ]
Derfus, Gayle [1 ]
Kluck, Brian [1 ]
Nogal, Bartek [1 ]
Emery, Craig [1 ]
Summers, Christie [1 ]
Zheng, Kai [1 ]
Bayer, Robert [1 ]
Amanullah, Ashraf [1 ]
Yan, Boxu [1 ]
机构：
[1] Genentech Inc, Dept Pharma Tech Dev, Oceanside, CA USA
来源：
MABS | 2011年 / 3卷 / 06期
关键词：
antibody; basic charge variants; proline amidation; copper; mass spectrometry; ALPHA-HYDROXYLATING MONOOXYGENASE; MONOCLONAL-ANTIBODY IGG1; POSTTRANSLATIONAL MODIFICATIONS; AMIDATION ACTIVITY; HETEROGENEITY;
D O I：
10.4161/mabs.3.6.17959
中图分类号：
R-3 [医学研究方法]; R3 [基础医学];
学科分类号：
1001 ;
摘要：
We report a case study of an IgG1 with a unique basic charge variant profile caused by C-terminal proline amidation on either one or two heavy chains. The proline amidation was sensitive to copper ion concentration in the production media during cell culture: the higher the Cu2+ ion concentration, the higher the level of proline amidation detected. This conclusion was supported by the analysis of samples that revealed direct correlation between the proline amidation level observed from peptide maps and the level of basic peaks measured by imaged capillary isoelectric focusing and a pH gradient ion-exchange chromatography method. The importance of these observations to therapeutic antibody production is discussed.
引用
收藏
页码：577 / 583
页数：7
相关论文
共 50 条
  • [1] Charge variants in IgG1 Isolation, characterization, in vitro binding properties and pharmacokinetics in rats
    Khawli, Leslie A.
    Goswami, Sirj
    Hutchinson, Ryan
    Kwong, Zephania W.
    Yang, Jihong
    Wang, Xiangdan
    Yao, Zhenling
    Sreedhara, Alavattam
    Cano, Tony
    Tesar, Devin
    Nijem, Ihsan
    Allison, David E.
    Wong, Pin Yee
    Kao, Yung-Hsiang
    Quan, Cynthia
    Joshi, Amita
    Harris, Reed J.
    Motchnik, Paul
    MABS, 2010, 2 (06) : 613 - 624
  • [2] Conformational characterization of the charge variants of a human IgG1 monoclonal antibody using H/D exchange mass spectrometry
    Tang, Liangjie
    Sundaram, Shanmuuga
    Zhang, Jingming
    Carlson, Ping
    Matathia, Alice
    Parekh, Babita
    Zhou, Qinwei
    Hsieh, Ming-Ching
    MABS, 2013, 5 (01) : 114 - 125
  • [3] BIOT 490-Identification and characterization of charge variants of a humanized IgG1 monoclonal antibody
    Vlasak, Josef
    Bussat, Marie C.
    Wang, Shiyi
    Wagner-Rousset, Elsa
    Schaefer, Mark
    Klinguer-Hamour, Christine
    Kirchmeier, Marc
    Corvaia, Nathalie
    Ionescu, Roxana
    Beck, Alain
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2008, 236
  • [4] Identification of multiple sources of the acidic charge variants in an IgG1 monoclonal antibody
    Shiwei Miao
    Panpan Xie
    Mao Zou
    Li Fan
    Xuping Liu
    Yan Zhou
    Liang Zhao
    Ding Ding
    Haibin Wang
    Wen-Song Tan
    Applied Microbiology and Biotechnology, 2017, 101 : 5627 - 5638
  • [5] Identification of multiple sources of the acidic charge variants in an IgG1 monoclonal antibody
    Miao, Shiwei
    Xie, Panpan
    Zou, Mao
    Fan, Li
    Liu, Xuping
    Zhou, Yan
    Zhao, Liang
    Ding, Ding
    Wang, Haibin
    Tan, Wen-Song
    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2017, 101 (14) : 5627 - 5638
  • [6] Characterization of acidic and basic variants of IgG1 therapeutic monoclonal antibodies based on non-denaturing IEF fractionation
    Dada, Oluwatosin O.
    Jaya, Nomalie
    Valliere-Douglass, John
    Salas-Solano, Oscar
    ELECTROPHORESIS, 2015, 36 (21) : 2695 - 2702
  • [7] Cysteine in cell culture media induces acidic IgG1 species by disrupting the disulfide bond network
    Prade, Elke
    Zeck, Anne
    Stiefel, Fabian
    Unsoeld, Andreas
    Mentrup, David
    Arango Gutierrez, Erik
    Gorr, Ingo H.
    BIOTECHNOLOGY AND BIOENGINEERING, 2021, 118 (03) : 1091 - 1104
  • [8] BIOT 159-Separation of charge variants of a humanized IgG1 monoclonal antibody
    Vlasak, Josef
    Green-Trexler, Erin
    Kirchmeier, Marc
    Ionescu, Roxana M.
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2008, 236
  • [9] Small conformational changes in IgG1 detected as acidic charge variants by cation exchange chromatography
    Fukuda, Masakazu
    Graewert, Melissa A.
    Jeffries, Cy M.
    Svergun, Dmitri I.
    Yamazaki, Tadao
    Koga, Akiko
    Yamanaka, Yuji
    ANALYTICAL BIOCHEMISTRY, 2023, 680
  • [10] Characterization of the Photodegradation of a Human IgG1 Monoclonal Antibody Formulated as a High-Concentration Liquid Dosage Form
    Qi, Pei
    Volkin, David B.
    Zhao, Hui
    Nedved, Michael L.
    Hughes, Robert
    Bass, Ryan
    Yi, Sun C.
    Panek, Mark E.
    Wang, Dana
    Dalmonte, Paul
    Bond, Michael D.
    JOURNAL OF PHARMACEUTICAL SCIENCES, 2009, 98 (09) : 3117 - 3130
12345