中文摘要：

目的探讨并建立表面等离子体免疫传感器芯片同步定量尿液中多种微量蛋白的方法。方法采用EDC/NHS方法将微量白蛋白（MA）、α-1微球蛋白（A1M）、β-2微球蛋白（B2M）、尿IgG的单克隆抗体固定于免疫传感器芯片之上,并用表面等离子体共振传感器（surface plasmon resonance,SPR）检测样本中微量蛋白的含量。利用4种微量蛋白的标准血清建立标准曲线,然后对其进行灵敏度、特异性等指标的方法学评价。以免疫散射比浊方法为参考方法,检测125例临床患者24 h尿液样本的微量蛋白浓度,比较2种方法检测相同样本时的结果差异。结果该表面等离子体共振免疫传感器芯片具有良好的操作性能和检测特异性。其标准曲线的R2均在0.98以上。其检测灵敏度分别是A1M为5μg/L,B2M为7.3μg/L,MA为11.5μg/L,IgG为22μg/L。所有4项指标可在20 min之内同步完成。与微量蛋白检测的经典方法免疫散射比浊法相比,两者的相关性和一致性较高。结论表面等离子体免疫传感器芯片可实现尿液中多种微量蛋白的快速、准确、同步定量。

英文摘要：

Objective To establish a novel methodology for simultaneous quantification of four trace proteins in urine with surface plasmon resonance（SPR） biosensor.Methods Monoclonal antibodies against Alpha-1 microglobulin（A1M）,Beta-2 microglobulin（B2M）,microalbumin（MA） and immunoglobulin G（IgG） were immobilized on the surface of biosensor chips,and then SPR was adopted to detect the corresponding antigens in standard solution containing the above-mentioned proteins.Calibrations of the 4 proteins were performed and the methodological evaluations were conducted to validate the sensitivity and specificity.Clinical urine samples from 125 inpatients were collected,and the contents of these 4 proteins were detected with SPR and immuno-turbidimetric assay at the same time.The results from the 2 methods were compared.Results The proposed SPR immunosensor had good signal-noise ratio and specificity.All the R2 of the calibration curves were higher than 0.98.The sensitivity of A1M was 5 μg/L,B2M 7.3 μg/L,MA 11.5 μg/L,and IgG 22 μg/L.All the detection was finished in 20 min.This proposed method had good consistency when compared with traditional immuno-turbidimetric assay.Conclusion This proposed SPR immunosensor chip can accurately and simultaneously determine multiple trace proteins in urine,and can be a useful alternative to current methodology.