Events including antibody-antigen affinity,internalization,trafficking and lysosomal proteolysis combinatorially determine the efficiency of antibody-drug conjugate(ADC) catabolism and hence the toxicity.Nevertheless,...Events including antibody-antigen affinity,internalization,trafficking and lysosomal proteolysis combinatorially determine the efficiency of antibody-drug conjugate(ADC) catabolism and hence the toxicity.Nevertheless,an approach that conveniently identifies proteins requisite for payload release and the ensuing toxicity for mechanistic studies and quality assessment is lacking.Considering the plethora of ADC candidates under development,we developed a target-responsive subcellular catabolism(TARSC) approach that examines ADC catabolism and probes changes in response to targeted interferences of proteins of interest.We firstly applied TARSC to study the commercial T-DM1 and the biosimilar.We recorded unequivocal catabolic behaviors regardless of the absence and presence of the targeted interferences.Their negligible differences in TARSC profiles agreed with their undifferentiated anti-tumoral efficacy according to further in vitro viability and in vivo tumor growth assays,highlighting TARSC analysis as a useful tool for biosimilarity assessment and functional dissection of proteins requisite for ADC catabolism.Additionally,we employed TARSC to investigate the catabolic behavior of a new trastuzumab-toxin conjugate.Collectively,TARSC can not only characterize ADC catabolism at(sub)cellular level but also comprehensively determine which protein targets affect payload release and therapeutic outcomes.Future use of TARSC is thus anticipated in early-stage screening,quality assessment and mechanistic investigations of ADCs.展开更多
基金financial support of the Natural Science Foundation of Jiangsu Province (BK20180079 and BK20180558, China)the Leading Technology Foundation Research Project of Jiangsu Province (BK20192005, China)+5 种基金the National Natural Science Foundation of China (82173783, 82173882, 81803625)Six Talent Peaks Project in Jiangsu Province (SWYY-101, China)International Industrial Technology Research Collaboration of Nanjing (201911008, China)the Innovative Research Groups of the National Nature Science Foundation of China (81421005)Nantong Science and Technology Project (JC2019133, JCZ18131, China)the Innovation Team of Affiliated Hospital of Nantong University (TFCT-A05, China)。
文摘Events including antibody-antigen affinity,internalization,trafficking and lysosomal proteolysis combinatorially determine the efficiency of antibody-drug conjugate(ADC) catabolism and hence the toxicity.Nevertheless,an approach that conveniently identifies proteins requisite for payload release and the ensuing toxicity for mechanistic studies and quality assessment is lacking.Considering the plethora of ADC candidates under development,we developed a target-responsive subcellular catabolism(TARSC) approach that examines ADC catabolism and probes changes in response to targeted interferences of proteins of interest.We firstly applied TARSC to study the commercial T-DM1 and the biosimilar.We recorded unequivocal catabolic behaviors regardless of the absence and presence of the targeted interferences.Their negligible differences in TARSC profiles agreed with their undifferentiated anti-tumoral efficacy according to further in vitro viability and in vivo tumor growth assays,highlighting TARSC analysis as a useful tool for biosimilarity assessment and functional dissection of proteins requisite for ADC catabolism.Additionally,we employed TARSC to investigate the catabolic behavior of a new trastuzumab-toxin conjugate.Collectively,TARSC can not only characterize ADC catabolism at(sub)cellular level but also comprehensively determine which protein targets affect payload release and therapeutic outcomes.Future use of TARSC is thus anticipated in early-stage screening,quality assessment and mechanistic investigations of ADCs.
