Study on the Reduction Conditions of Interchain Disulfide Bonds of IgG1 Antibody
Antibody-drug conjugates(ADCs)are a novel targeted cancer treatment composed of an antibody,payload,and linker.Reducing interchain disulfide bonds in the antibody to free thiols and subsequent Michael addition with the electrophilic moiety on the linker is a crucial method for constructing ADCs.This study meticulously explores the influence of three primary variables on the efficiency of reducing interchain disulfide bonds in antibodies:The molar equivalent ratio of the reducing agent to the monoclonal antibody,the temperature of the reaction,and the duration of the reaction.Additionally,the study involves the synthesis of drug com-pounds containing maleimide and a cytotoxic payload,which are then conjugated with reduced monoclonal antibodies to create ADC molecules.The Drug-to-Antibody Ratio(DAR)of these synthesized ADCs is subsequently assessed.Experimental data from this study reveals a clear relationship between the amount of reducing agent used and the reduction of interchain disulfide bonds within the antibodies.Notably,the reduction efficiency remains relatively consistent when the reducing agent dosage ranges from 2-fold to 10-fold molar equivalents to the antibody,but a significant variation is observed beyond 10-fold molar equivalents.Optimal reduction is achieved within a 4-hour timeframe when the reducing agent dosage is at 50-fold molar equivalents,with the reaction exhibiting peak efficiency at a temperature of 37 ℃.The study concludes that the most effective reduction conditions for antibodies entail a reducing agent dosage of 50-fold molar equivalents,a reaction temperature of 37 ℃,and completion within a 4-hour timeframe.More-over,the conjugation efficiency of the synthesized molecules containing maleimide and the cytotoxic payload to antibodies is reported to be an impressive 83.3％.These findings not only lay a robust scientific foundation for the production of ADCs but also offer valuable insights for future research and practical applications in the field of targeted cancer therapy.
NETL
NSTL
万方数据
