Enzyme-linked immunosorbent assays for quantification of MMMAE-conjugated ADCs and total antibodies in cynomolgus monkey sera
- Author:
Pei MIN
1
,
2
;
Liu TINGTING
;
Ouyang LU
;
Sun JIANHUA
;
Deng XIAOJIE
;
Sun XIAOMIN
;
Wu WEI
;
Huang PENG
;
Chen YI-LI
;
Tan XIAORONG
;
Liu XIAOYUE
;
Zhu PENG
;
Liu YONGZHEN
;
Wang DEHENG
;
Wu JUNLIANG
;
Wang QI
;
Wang GUIFENG
;
Gong LIKUN
;
Qin QIUPING
;
Wang CHUNHE
Author Information
1. Biotherapeutics Discovery Research Center,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai,201203,China
2. School of Pharmacy,University of Chinese Academy of Sciences,Beijing,100049,China
- Keywords:
Monomethyl auristatin E;
Antibody-drug conjugates;
Pharmacokinetics;
Trophoblast cell surface antigen 2
- From:
Journal of Pharmaceutical Analysis
2022;12(4):645-652
- CountryChina
- Language:Chinese
-
Abstract:
Antibody-drug conjugates(ADCs)are commonly heterogeneous and require extensive assessment of exposure-efficacy and exposure-safety relationships in preclinical and clinical studies.In this study,we report the generation of a monoclonal antibody against monomethyl auristatin E(MMAE)and the development,validation,and application of sensitive and high-throughput enzyme-linked immunosor-bent assays(ELISA)to measure the concentrations of MMAE-conjugated ADCs and total antibodies(tAb,antibodies in ADC plus unconjugated antibodies)in cynomolgus monkey sera.These assays were suc-cessfully applied to in vitro plasma stability and pharmacokinetic(PK)studies of SMADC001,an MMAE-conjugated ADC against trophoblast cell surface antigen 2(TROP-2).The plasma stability of SMADC001 was better than that of similar ADCs coupled with PEG4-Val-Cit,Lys(m-dPEG24)-Cit,and Val-Cit linkers.The developed ELISA methods for the calibration standards of ADC and tAb revealed a correlation be-tween serum concentrations and the OD450 values,with R2 at 1.000,and the dynamic range was 0.3-35.0 ng/mL and 0.2-22.0 ng/mL,respectively;the intra-and inter-assay accuracy bias%ranged from-12.2%to-5.2%,precision ranged from-12.4%to-1.4%,and the relative standard deviation(RSD)was less than 6.6%and 8.7%,respectively.The total error was less than 20.4%.The development and validation steps of these two assays met the acceptance criteria for all addressed validation parameters,which suggested that these can be applied to quantify MMAE-conjugated ADCs,as well as in PK studies.Furthermore,these assays can be easily adopted for development of other similar immunoassays.