Enzyme-linked immunosorbent assays for quantification of MMAE-conjugated ADCs and total antibodies in cynomolgus monkey sera  

作　　者：Min Pei Tingting Liu Lu Ouyang Jianhua Sun Xiaojie Deng Xiaomin Sun Wei Wu Peng Huang Yi-Li Chen Xiaorong Tan Xiaoyue Liu Peng Zhu Yongzhen Liu Deheng Wang Junliang Wu Qi Wang Guifeng Wang Likun Gong Qiuping Qin Chunhe Wang 

机构地区：[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 [3]Center for Drug Safety Evaluation and Research,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai,201203,China [4]Research and Development Center,Dartsbio Pharmaceuticals Ltd.,Zhongshan,Guangdong,528400,China [5]Zhongshan Institute for Drug Discovery,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Zhongshan,Guangdong,528400,China [6]School of Pharmacy,Shanghai Engineering Research Center of Immunotherapeutics,Fudan University,Shanghai,201203,China

出　　处：《Journal of Pharmaceutical Analysis》2022年第4期645-652,共8页药物分析学报（英文版）

基　　金：supported by the China National Major Scientific and Technological Special Project for“Significant New Drugs Innovation and Development”(Grant No.:2019ZX09732002-006);the National New Drug Creation Program of China(Grant No.:2018ZX09201017-004);the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant Nos.:XDA12020223,XDA12020330,XDA12020360,and XDA12050305);the National Natural Science Foundation of China(Grant Nos.:81872785 and 81673347);the Science and Technology Planning Projects of Department of Science and Technology Province(Grant No.:20190202);Shanghai Municipal Commission of Science and Technology of China(Grant Nos.:17431904400,19YF1457400,and 21S11904500);Institutes for Drug Discovery and Development,the Chinese Academy of Sciences(Grant Nos.:CASIMM0120202007 and CASIMM0120202008);Major Scientific and Technological Special Project of Zhongshan City(Grant Nos.:191022172638719 and 210205143867019).

摘　　要：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 immunosorbent 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 successfully 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 between 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.

关 键 词：Monomethyl auristatin E Antibody-drug conjugates PHARMACOKINETICS Trophoblast cell surface antigen 2 

分 类 号：R969[医药卫生—药理学]