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Development and Validation of a Simoa Assay for Determination of Recombinant Batroxobin in Human Serum

Current Medical Science 2021年第3期41卷 618-625页

作者： Bian-zhen WANG Meng-jia WANG Min-rui WANG Lun OU Li-hou DONG Kelly DONG Hai-feng SONG State Key Laboratory of Proteomics Beijing Proteome Research CenterNational Center of Protein Sciences(Beijing)Beijing Institute of LifeomicsBeijing 102206China United-Power Pharma Tech Co LtdBeijing J02206China

Recombinant batroxobin(S3101)is a thrombin-like serine protease that binds to fibrinogen or is taken up by the reticuloendothelial system.A literature survey showed no adequate method that could determine sufficient concentrations to evaluate pharmacokinetic parameters for phase I clinical ***,a sensitive method is urgently needed to support the clinical pharmacokinetic evaluation of *** this study,a sensitive bioanalytical method was developed and validated,using a quanterix single molecular array(Simoa)***,to thoroughly assess the platform,enzyme-linked immunosorbent assay and electrochemiluminescence assay were also developed,and their performance was compared with that of this novel technology *** assay was validated in compliance with the current *** with the Simoa assay were precise and accurate,presenting a valid assay range from 6.55 to 4000 pg/*** intra-and inter-run accuracy and precision were within-19.3%to 15.3%and 5.5%to 17.0%,respectively.S3101 was stable in human serum for 280 days at-20℃and-70℃,for 2 h prior to pre-treatment and 24 h post pre-treatment at room temperature(22℃-28℃),respectively,and after five and two freeze-thaw cycles at-70℃and-20oC,*** Simoa assay also demonstrated sufficient dilution linearity,assay sensitivity,and parallelism for quantifying S3101 in human *** Simoa assay is a sensitive and adequate method for evaluating the pharmacokinetic parameters of S3101 in human serum.

关键词： enzyme-linked immunosorbent assay electrochemiluminescence assay quanterix single molecular array recombinant batroxobin ligand-binding assay

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