LC-MS analysis of sintilimab as an anti-PD-1 therapeutic mab for its improved hinge stability study
10.16438/j.0513-4870.2018-0574
- VernacularTitle:LC-MS法对抗PD-1信迪利单抗药物分子铰链区序列改构稳定性的研究
- Author:
Chuan-fei YU
1
;
Xing-jun CAO
2
;
Wen-bo WANG
1
;
Hai-qing NI
2
;
Amy Y GUO
;
Lan WANG
1
Author Information
1. National Institutes for Food and Drug Control, Beijing 102629, China
2. Innovent Biologics Suzhou Co. Ltd, Suzhou 215123, China
- Publication Type:Research Article
- Keywords:
IgG4;
Fab-arm exchange;
LC-MS;
PD-1;
sintilimab
- From:
Acta Pharmaceutica Sinica
2019;54(1):122-129
- CountryChina
- Language:Chinese
-
Abstract:
Monoclonal antibodies (mAbs) have been widely used as therapeutic drugs for treating diseases such as cancers and auto-immune diseases. When using an IgG4 isotype, one of the challenges is the instability of its hinge which is prone to Fab-arm exchange (FAE). The hinge sequence of a wild type IgG4 is -CPSC-, however, a single point mutation S228P from -CPSC- to -CPPC- can effectively diminish FAE, thereby improving hinge stability of the IgG4 molecule. Sintilimab is the fully human anti-PD-1 monoclonal antibody designed and developed for immuno-oncology, in which serine 228 in the hinge was engineered to proline to mitigate FAE. In this study, LC-MS is used to study hinge stability of sintilimab in both in vitro (PBS and human serum) and in vivo (SCID mouse) studies. The studies demonstrate that LC-MS is a fast and simple way to monitor for the occurrence of FAE in vitro and in vivo, and FAE can be eliminated by antibody engineering with a single point mutation.
