Gogolin K J, Slaughter C A, Harris H
Proc Natl Acad Sci U S A. 1981 Aug;78(8):5061-5. doi: 10.1073/pnas.78.8.5061.
Enzyme--monoclonal antibody complexes formed between six different monoclonal antibodies and the six phenotypes of human placental alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] that represent the homozygous and heterozygous combinations of the three common alleles have been examined by electrophoresis in starch, acrylamide, and agarose gels. Since the complexes formed retain full enzyme activity, they could be detected after gel electrophoresis by an enzyme stain. Distinctive electrophoretic patterns were obtained with each monoclonal antibody. Differential binding of certain of the antibodies with the products of different alleles produces clear discrimination of various homozygous and heterozygous phenotypes. This discrimination parallels the results previously obtained by using a quantitative binding radioimmunoassay. The results show that this general method should prove useful in screening hybridoma fluids for the presence of monoclonal antibodies to specific enzymes; in the detection of allelic variation, even where this is not expressed by electrophoretic differences among the uncomplexed enzymes; and in discriminating between homozygotes and heterozygotes. It could also prove to be a useful tool in the elucidation of the molecular structures of enzyme--monoclonal antibody complexes.
六种不同的单克隆抗体与人类胎盘碱性磷酸酶[正磷酸单酯磷酸水解酶（最适pH为碱性），EC 3.1.3.1]的六种表型形成的酶-单克隆抗体复合物已在淀粉凝胶、丙烯酰胺凝胶和琼脂糖凝胶中进行了电泳分析，这六种表型代表了三个常见等位基因的纯合和杂合组合。由于形成的复合物保留了全部酶活性，因此在凝胶电泳后可用酶染色法进行检测。每种单克隆抗体都获得了独特的电泳图谱。某些抗体与不同等位基因产物的差异结合能够清晰地区分各种纯合和杂合表型。这种区分与先前使用定量结合放射免疫测定法获得的结果相似。结果表明，这种通用方法在筛选杂交瘤细胞培养液中是否存在针对特定酶的单克隆抗体、检测等位基因变异（即使未复合的酶之间没有电泳差异表现）以及区分纯合子和杂合子方面应会被证明是有用的。它也可能被证明是阐明酶-单克隆抗体复合物分子结构的有用工具。
