摘要
为建立特异有效的脂蛋白（ａ）检测方法，应用杂交瘤技术，制备出４株抗载脂蛋白（ａ）单克隆抗体，分别命名为Ｍ＿１、Ｍ＿２、Ｍ＿３、Ｍ＿４。各株单克隆抗体与纤溶酶原和载脂蛋白Ｂ等无交叉反应。单抗相加试验及竞争抑制试验结果表明：Ｍ＿１、Ｍ＿４识别同一抗原位点，Ｍ＿２、Ｍ＿３识别另一位点。选用Ｍ＿１、Ｍ＿２、Ｍ＿４３株单克隆抗体建立了测定血清脂蛋白（ａ）浓度的双抗体夹心酶联免疫测定法。样品稀释后测定范围为０．０５～１．６０ｍｇ／Ｌ，批内平均变异系数（ＣＶ）４．２％，批间平均ＣＶ８．７％。测定了１０３２例中老年临床血清标本，脂蛋白（ａ）浓度为１７９，１±１７９．３ｍｇ／Ｌ（ｘ±ｓ）。与多克隆抗体的酶联免疫测定法同时测定了３８份临床标本，结果相似。此法可满足临床脂蛋白（３）浓度检测的需要。
our clones of hybridoma which secreted monoclonal antibodies named as M_1、M_2、M_3 and M_4 to apolipoprotein(a)were developed.These monoclonal antibodies showed no cross-reaction to apolipopro-tein B and plasminogen. Immunologically,the M_1 and M_4 recognized the same antigenic epitope,where-as M_2 and M_3 recognized another. A sandwich enzyme-linked immunosorbent assay was established for determining serum lipoprotein(a)concentration with mixed McAbs (M_1、M_2 and M_4).The quantitative range is 0. 05～1. 60mg/L. The average intra-and inter-assay coefficient of variation were 4.2%and 8.7%respectively. The sera concentration of Lp(a ) in 103 2 middle-aged and old patients were mea-sured,and the result was 179.1±179.3mg/L.The results of this assay corresponded well to that of the ELISA method using polycolonal antibody to apolipoprotein(a).We concluded that this method was suitable for routine usage in clinical laboratories.
