A method for measuring the ADCP potency of anti-CD38 monoclonal antibody
10.16438/j.0513-4870.2021-1613
- VernacularTitle:利用实验设计优化和建立抗CD38单克隆抗体ADCP生物学活性测定方法
- Author:
Chun-yu LIU
1
;
Chuan-fei YU
1
;
Xin LI
2
;
Zhi-hao FU
1
;
Yong-fei CUI
1
;
Lu-yun GUO
1
;
Lan WANG
1
Author Information
1. Division of Monoclonal Antibody, National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, Beijing 102629, China
2. Guangzhou Insititue for Drug Control, Guangzhou 510160, China
- Publication Type:Research Article
- Keywords:
anti-CD38 monoclonal antibody;
antibody-dependent cell-mediated phagocytosis;
biological activity;
esign of experiment;
Plackett-Burman design;
optimization;
validation
- From:
Acta Pharmaceutica Sinica
2022;57(5):1459-1464
- CountryChina
- Language:Chinese
-
Abstract:
A method to measure the antibody-dependent cell-mediated phagocytosis (ADCP) potency of anti-CD38 mAb was developed based on design of experiment (DoE) with a Jurkat/NFAT/CD32a-FcεRIγ transgenic cell line as the effector cell, the Daudi cell line as the target cells, and luciferase as the detection system. The DoE method was used for optimization of experimental parameters and methodological validation. The results show that anti-CD38 mAb exhibits a dose-response relationship with the following four-parameter equation: y = (A - D) / [1 + (x / C)B] + D. Several experimental parameters were optimized by statistical experimental design and determined as follows: the working concentration of anti-CD38 mAb was 800-20.81 ng·mL-1, the density of the target cells was 7.5×104 per well, and the density of effector cells was 2.5×104 per well, with an induction time of 6 h. The method showed good specificity. The recovery rate for samples from 5 different groups showed that the relative potencies of anti-CD38 mAb were (59.97 ± 4.74) %, (82.44 ± 5.15) %, (110.69 ± 11.71) %, (129.23 ± 5.22)% and (162.15 ± 3.66) %. The recoveries ranged from 103% to 120% and the RSDs of the above results were all less than 11%. The linear detection range was 50%-150%. Based on DoE design, this method for measuring ADCP potency of anti-CD38 mAb was optimized and validated with good specificity, repeatability and accuracy. This method can be used for evaluation of ADCP biological activity of anti-CD38 mAbs.
