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
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
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