Size heterogeneity analysis of monoclonal antibody products
10.3760/cma.j.issn.0254-5101.2014.09.013
- VernacularTitle:人源化抗 VEGF 单克隆抗体制品的大小异质性分析
- Author:
Chuanfei YU
;
Wenbo WANG
;
Meng LI
;
Lan WANG
;
Feng ZHANG
;
Chunyu LIU
;
Kai GAO
- Publication Type:Journal Article
- Keywords:
Monoclonal antibody;
Size heterogeneity;
CE-SDS;
SE-HPLC;
Aggregate;
Fragment
- From:Chinese Journal of Microbiology and Immunology
2014;(9):718-722
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the capability of capillary electrophoresis-sodium dodecyl sul-fate ( CE-SDS) and size exclusion-high performance liquid chromatography ( SE-HPLC) for analysis of size heterogeneity of monoclonal antibody products .Methods The size heterogeneity of one humanized anti-VEGF monoclonal antibody was analyzed by using non-reduced and reduced CE-SDS, and conventional , de-natured and denatured reduced SE-HPLC.Results The percentage of aggregates detected by non-reduced CE-SDS (0.82%±0.01%) was equal to that by using denatured SE-HPLC (1.05%±0.02%), but it was significantly lower than that by using conventional SE-HPLC analysis (5.08%±0.10%).With regard to fragments analyzed with non-reduced antibodies, its percentage was (7.12±0.04)% measured by non-re-duced CE-SDS analysis that was significantly higher than that by conventional SE -HPLC analysis (0.02%± 0.01%) and denatured SE-HPLC analysis (0.62%±0.01%).Using reduced antibodies , the percentage of fragments was (3.19±0.50)%tested by reduced CE-SDS analysis that was significantly higher than that by using denatured reduced SE-HPLC analysis (0.07%±0.01%).Conclusion Conventional SE-HPLC was more objective than CE-SDS for content analysis of aggregates , as both the covalent and non-covalent forms of aggregates could be detected .Non-reduced CE-SDS could demonstrate the content of clips , while reduced CE-SDS showed the degraded fragments .Therefore, CE-SDS had an advantage over conventional SE-HPLC for content analysis of fragments .The use of the two analytical methods in combination provided solid techni-cal supports for the quality control of size heterogeneity of monoclonal antibodies .
