Objective To establish an enzyme-linked immunoassay for the detection of single-chain precursor residues in insulin glargine so as to recommend a new method for quality control and safety evaluation of insulin glargine.Methods An enzyme-linked immunoassay based on double antibody sandwich was established by using proinsulin glargine as the standard.The repeatability,accuracy,scope and precision of the method were verified,and nine batches of insulin glargine drug substances were tested using this method.Results Four-parameter curve fitting was performed using the proinsulin glargine standard at concentrations ranging from 0.156 to 10.00 ng·mL-1,and the correlation coefficient of the curve exceeded 0.99.This method was highly accurate and repeatable.Three batches of insulin glargine drug substances were repeatedly detected.The average recovery ranged from 104%to 113%,and the RSD%was less than 10%,suggesting the good precision of this method.The single-chain precursor residues in all the nine batches were less than 1 ng·mg-1.Conclusion The enzyme-linked immunoassay established in this study can be used for quality control and safety evaluation of single-chain precursor residues in insulin glargine and other recombinant human insulin analogues.
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