重组人血清白蛋白抗体检测方法的建立及确证

目的：建立一种高灵敏度、高特异性、操作简单快捷、通量高的重组人血清白蛋白（rHSA）抗体检测方法。方法：采用桥连ELISA法，即将rHSA包被于96孔板，加入待测血样及阳性对照，用辣根过氧化物酶标记的rHSA检测，显色读取D_450nm/D_570nm值：用此方法确定临界值、方法灵敏度、精密度、血药浓度对检测方法的影响，再以免疫清除法进行确证。结果：通过桥连ELISA法确定临界值为0．0492，方法灵敏度为352ng／mL，方法板间、板内精密度均小于20％，且血药中的rHSA浓度为20μg／mL时不影响抗体的检测；经免疫清除法可将假阳性样本排除，从而提高了方法的特异性．结论：建立的方法可以准确、快速地检测出rHSA的特异性抗体。

Establishment and Confirmed of Recombinant Human Serum Albumin Antibody Detection Method

Objective： To develop a high specific, high sensitive bridging ELISA method for the quantification of recombinant human serum albumin（rHSA） antibody. Methods： The microplate was coated with rHSA, and a HRP-labeled rHSA was used as the detection antibody for the bridging ELISA. The established bridging ELISA would be evaluated by the cut off, sensitivity, precision, plasma concentration to the influence of method. Use immune clearance to prove the method convincingly. Results： The cut off value was 0.0492, the sensitivity of the bridging ELISA with a limit of detection was 352 ng/mL, the intra- and inter-assay precisions were less than 20%, and the plasma concentration was 20 μg/mL that didn＇t influence the detection of antibody. The immune clearance could exclude false positive samples. Conclusion： This method is accurate, fast for the determination of rHSA antibody.