Chiplunkar S V, Amin K M, Gangal S G
Indian J Med Res. 1989 Feb;90:1-8.
Four anti-alphafoetoprotein (AFP) monoclonal antibodies (MAb) were raised in the laboratory and characterized using ELISA and immunodot assays. The affinity constants of the MAbs, analysed by scatchard plots, ranged from 3.1 X 10(8) to 2.15 X 10(9) M/l. Epitope analysis using competition RIA indicated that MAb 5E2D7 and 5E2E3 recognize different epitopes on AFP. This combination was used to set up a two site sandwich ELISA with HRPO conjugated 5E2D7. AFP values in sera of hepatocellular carcinoma patients and pregnant women were quantitated using sandwich ELISA. The anti-AFP MAbs showed strong reactivity when tested on hepatoma tissue sections using immunoperoxidase technique.
在实验室中制备了四种抗甲胎蛋白(AFP)单克隆抗体(MAb),并使用酶联免疫吸附测定(ELISA)和免疫斑点测定对其进行了表征。通过Scatchard图分析,这些单克隆抗体的亲和常数范围为3.1×10⁸至2.15×10⁹M/l。使用竞争放射免疫分析(RIA)进行的表位分析表明,单克隆抗体5E2D7和5E2E3识别AFP上不同的表位。利用这种组合,用辣根过氧化物酶(HRPO)偶联的5E2D7建立了双位点夹心ELISA。使用夹心ELISA对肝细胞癌患者和孕妇血清中的AFP值进行了定量。当使用免疫过氧化物酶技术在肝癌组织切片上进行检测时,抗AFP单克隆抗体表现出强烈的反应性。