HPA-1a抗体SPR技术定量检测方法的建立

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,...

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

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.