脂蛋白(α)单克隆抗体对抗原决定簇的测定及应用研究

Determination of Antigen Determinants by Monoclonal Antibodies to Human Applipoprotein (α)

应田淋巴细胞杂交瘤技术,建立了4株抗人血清脂蛋白(α)(LP(a))杂交瘤细胞株(BH6、BH7、BH11和BH21),并以制备的单克隆抗体(McAb)对其抗原决定簇及免疫学特性进行了分析。Ig亚类测定:BH6为IgG1,BH7、BH11和BH21均为IgG2a,腹水效价为1×10^(-7)。特异性测定:LP(α)McAb与载脂蛋白(apo)AⅠ、AⅡ、B、CⅠ、CⅡ、CⅢ、E,人血清白蛋白,纤维蛋白溶酶原(pg)没有交叉反应。单抗相加试验和双抗体竞争试验结果证实,BH6和BH21为识别LP(α)上同一抗原决定簇的McAb;BH7和BH11则为针对LP(α)上另一抗原决定簇的McAb。分别利用单株和混合株McAb标记酶建立了可应用于人血清LP(α)含量测定的ELISA双抗体夹心法。

Four hybridoma cell lines secreting monoclonal antibodies (McAbs ) to human serum apolipoprotien ( a ) were produced by lymphocyte hybridoma technique. These McAbs were designated BH6, BH7, BH11 and BH21 and the class and subclass of these immunoglobulins were determined. BH6 was identified as IgG1, BH7, BH11 and BH21 were IgG2a. The antibody titer of the ascitic fluids was 1×10-7. Antibody specificity analysis showed that these McAbs to apo ( a ) did not cross react with apolipop- rotein AⅠ, AⅡ, B, CⅠ, CⅡ, CⅢ, E, human serum albumin and piasminogen. The results of cotitration test and competitive antibody binding assay showed that McAbs BH6 and BH21 recognized the same antigen determinant and McAbs BH7 and BH11 also recognized an identical antigen determinant. Sandwich ELLSAs for determining human sera LP (α) concentration were established with a single McAb and mixed McAbs respectively.