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.

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

Biotherapeutics Discovery Research Center,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai,201203,China;School of Pharmacy,University of Chinese Academy of Sciences,Beijing,100049,China;Center for Drug Safety Evaluation and Research,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai,201203,China;Research and Development Center,Dartsbio Pharmaceuticals Ltd.,Zhongshan,Guangdong,528400,China;Zhongshan Institute for Drug Discovery,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Zhongshan,Guangdong,528400,China;School of Pharmacy,Shanghai Engineering Research Center of Immunotherapeutics,Fudan University,Shanghai,201203,China

药物分析学报（英文）

[1]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-.Enzyme-linked immunosorbent assays for quantification of MMMAE-conjugated ADCs and total antibodies in cynomolgus monkey sera)[J].药物分析学报（英文）,2022(04):645-652

MMMAE,auristatin,tAb,SMADC001,TROP,PEG4,dPEG24

Enzyme,linked,immunosorbent,quantification,ADCs,total,antibodies,cynomolgus,monkey,sera,Antibody,drug,conjugates,are,commonly,heterogeneous,require,extensive,assessment,exposure,efficacy,safety,relationships,preclinical,studies,In,this,study,report,generation,monoclonal,antibody,against,monomethyl,development,validation,application,sensitive,high,throughput,enzyme,measure,concentrations,plus,unconjugated,These,were,suc,cessfully,applied,vitro,plasma,stability,pharmacokinetic,PK,trophoblast,cell,surface,antigen,was,better,than,that,similar,coupled,Val,Cit,Lys,linkers,developed,methods,calibration,standards,revealed,correlation,tween,serum,OD450,values,dynamic,respectively,intra,inter,accuracy,bias,ranged,from,precision,relative,deviation,RSD,less,error,steps,these,two,acceptance,criteria,all,addressed,parameters,which,suggested,can,quantify,well,Furthermore,easily,adopted,other,immunoassays

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