抗程序性死亡配体-1单克隆抗体在食蟹猴中单个核细胞的受体占位方法学比较

Comparison of receptor occupancy methods of anti-programmed death ligand 1 monoclonal antibody inmononuclear cell of cynomolgus monkey

目的比较抗程序性死亡配体-1(programmed death ligand 1,PD-L1)抗体在食蟹猴全血与外周血单个核细胞(peripheral blood mononuclear cell,PBMC)2种样本类型中的受体占位方法学。方法分别建立了基于流式细胞术的食蟹猴全血与PBMC中抗PD-L1单克隆抗体受体占位方法,并对2种样本中建立的方法进行验证对比。结果全血的批内和批间精密度分别是2.4%~24.4%和11.4%~12.9%,全血采集后24 h内稳定,样本固定后24 h内稳定。PBMC的批内和批间精密度分别是1.3%~11.5%和4.8%~8.3%,全血采集后24 h内稳定,固定后24 h内稳定,PBMC冻存3个月内稳定。2种样本的选择性、特异性和灵敏度无显著差别。结论多中心临床试验时为保证样本的及时处理及降低物流成本,PBMC可替代全血作为受体占位实验样本。

Objective To compare receptor occupancymethods of anti-programmed death ligand 1(anti-PD-L1)antibodyin whole blood of cynomolgus monkey and peripheral blood mononuclear cell(PBMC).Methods Receptor occupancy(RO)methods based on flow cytometry for anti-PD-L1 monoclonal antibody in cynomolgus monkey whole blood and PBMC were established,and the RO methods established in the two samples were comprehensively verified.Results The intra-run and inter-run precisions of the whole blood were 2.4%to 24.4%and 11.4%to 12.9%.The whole blood was stable within 24 hours after collection and the sample was stable within 24 hours after fixation.The intra-and inter-run precisions of the PBMC were 1.3%to 11.5%and 4.8%to 8.3%.It was stable within 24 hours after whole blood collection and stable within 24 hours after fixation,and PBMC was stable within 3 months after freezing.There were no significant differences in selectivity,specificity,and sensitivity between the two samples.Conclusion Cynomolgus monkey PBMC could replace whole blood as the receptor occupancy sample in multi-center clinical trials to ensure the timely processing of samples and reduce logistics costs.