人血清载脂蛋白AⅡ酶联免疫测定法研究

ELISA for Measurement of Human Serum Apolipoprotein A Ⅱ

本文介绍测定人血清载脂蛋白ＡⅡ（ａｐｏＡⅡ）的酶联免疫法（ＥＬＩＳＡ）。特异抗人ａｐｏＡⅡ血清由免疫山羊获得，按过碘酸钠法制备辣根过氧化物酶羊抗人ａｐｏＡⅡγ球蛋白交联物。用羊抗人ａｐｏＡⅡγ球蛋白（２０μｇ／ｍｌ）包被聚苯乙烯酶标板。结果表明：ａｐｏＡⅡ抗血清与ａｐｏＡⅠ、ａｐｏＣｓ，ａｎｏＢ１００（ＬＤＬ）以及人血清白蛋白无交叉反应，最小检测量为５０Ｏｎｇ，标准曲线工作范围为０．２５～８．００ｍｇ／ｄｌ，板内及板间变异系数分别为５．０％～９．２％及６．８％～１１．７％，回收率１０６．０±２．１％（ｎ＝４）。应用本法测得４１例血脂正常者ａｐｏＡⅡ含量为２４．４±５．９ｍｇ／ｄｌ，与用ＲＩＤ法测得的结果２６．７±４．６ｍｇ／ｄｌ相比（ｒ＝０．８０００，Ｐ＜０．００１），无显著性差异。本法特异性强，灵敏度高，简便快速。

A specific,sensitive and simple ELISAfor the measurement of human serum apo A Ⅱ has beendeveloped. The monospecific antibody was raised ingoats. The polytyrene plates coated with purified anti-apo A Ⅱ goat γ-globulin together with enzyme labelledgoat antibodies against human apo A Ⅱ conjuate wereused in this assay。 The conjuate was obtained by bind-ing horseradish perioidate by a simplified periedasemthod。 No cross-reactivity with human apo A I,B100.C Ⅰ.C Ⅱ,C Ⅲ and albumin was observed.Theminimum measurable concantration of apo A Ⅱ was500ng in each assay. A standard curve with a workingrange of 0.25-8.0 mg/dl was plotted. The coefficientsof va riation of the repreducibility of intra-and inter-assays of apo A Ⅱ in samples were 5.0-3.6% and 6.8-9.9% respectively,The recovery were 106. 0±2.1%(n= 4 ).The mean concentrations of apo A Ⅱ in 41healthy subjects were 24.4±15.9mg/dl by our method and 26.7±14.6mg/dl by RID method ,respectively(r=o.8000·P<0.001).