HPA-1a抗体SPR技术定量检测方法的建立  

Establishment of Quantitative SPR Assay for Antibodies Against Human Platelet Antigen-1a

作　　者：李睿书[1] 倪铭晨 朱慧君[1] 马勤勤 傅敏[1] 陆萍[1] LI Rui-Shu;NI Ming-Chen;ZHU Hui-Jun;MA Qin-Qin;FU Min;LU Ping(Shanghai Blood Center,Shanghai Institute of Blood Transfusion,Shanghai 200051,China;Department of Laboratory Medicine,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]上海市血液中心上海市输血研究所,上海200051 [2]上海交通大学医学院附属瑞金医院检验系,上海200025

出　　处：《中国实验血液学杂志》2021年第1期239-242,共4页Journal of Experimental Hematology

摘　　要：目的:建立针对人类血小板抗原(HPA)-1a抗体的表面等离子体共振(SPR)技术定量检测方法。方法:以氨基偶联法在芯片表面偶联重组蛋白,采用SPR方法检测不同浓度的标准抗血浆样本,优化实验条件,在检测范围内绘制标准曲线,并分析其敏感性、特异性、准确性和精密度。结果:利用SPR技术建立针对HPA-1a抗体的定量检测方法,检测范围为0-20 IU,准确性(回收率)为97.75%-103.08%,批内精密度[变异系数(CV)]为3.53%-4.29%;批间精密度(CV)为2.08%-4.40%。特异性实验中,以不同的血小板膜蛋白单抗和含相应抗体的参比血浆进行检测,均未见明显阳性结果。结论:建立的针对HPA-1a抗体的SPR技术定量检测方法具有较高的敏感性、特异性、准确性和精密度,且操作简便快速,这为科研和临床相关抗体的检测提供了一种新的参考方法。Objective:To establish quantitative surface plasmon resonance(SPR) assay for antibodies against human platelet antigen-1 a(HPA-1 a).Methods:Recombinant protein was fixed on the chip surface by amino coupling method.SPR assay was used to detect the standard antibodies against HP A-1 a at different conceatration.The optimal experimental parameters were determined,and standard curves were constructed with linear regression.Moreover,the sensitivity,specificity,accuracy and precision of the assay were evaluated.Results:The quantitative SPR assay for HPA-la antibodies was established.The determination ranges were 0-20 IU,with accuracy(recovery rate) was 97.75%-103.08%.The intraassay precision [coefficients of variation(CV)] was 3.53%-4.29%,and the inter-assay precision(CV) was 2.08%-4.40%.For specificity test,several kinds of monoclonal and human antibodies against platelet membrane protein were tested and no positive result was observed.Conclusion:The established quantitative SPR assay for HPA-la antibodies shows good sensitivity,specificity,accuracy and precision,and this rapid and simple method provides a new reference method for scientific research and clinical antibody detection.

关 键 词：人类血小板抗原 表面等离子共振 定量 安全输血 

分 类 号：R457.1[医药卫生—治疗学]