Slaughter C A, Gogolin K J, Coseo M C, Meyer L J, Lesko J, Harris H
Am J Hum Genet. 1983 Jan;35(1):1-20.
Eighteen monoclonal antibodies were produced by the mouse hybridoma method using purified placental alkaline phosphatase (ALP) as antigen. The ability of the various antibodies to discriminate among allelic variants of the enzyme was tested using a large panel of placental ALPs that had been typed electrophoretically. The panel included sets of samples of each of the six common polymorphic phenotypes as well as a series of rare variants. The reactivity of each antibody with each placental ALP (binding ratio) was determined relative to a single standard placental ALP (type 1) in a quantitative binding assay. The findings for six of the antibodies have already been reported. The results on the other 12 antibodies are presented here, and the combined data on the total series of 18 antibodies are analyzed and discussed. Six of the 18 antibodies showed significantly reduced binding to one or another of the products of the three common alleles. In three cases, the discrimination was reflected by essentially "all-or-none" binding reactions. In the other three cases, the binding differences were less marked but could be demonstrated by quantitative comparisons of the binding ratios. Quantitative binding ratio comparisons also enabled heterozygotes to be differentiated from homozygotes in each case. Some of the antibodies showed reduced binding with certain of the rare variant ALP electrophoretic phenotypes. It is estimated that at a minimum this unselected series of 18 antibodies is directed to at least nine different antigenic determinants on the surface of the placental ALP molecule. The results illustrate the power of monoclonal antibodies to discriminate among allelic variants of enzymes.
以纯化的胎盘碱性磷酸酶（ALP）作为抗原，通过小鼠杂交瘤法制备了18种单克隆抗体。使用大量经电泳分型的胎盘ALP对各种抗体区分该酶等位基因变体的能力进行了测试。该样本库包括六种常见多态性表型中每种表型的样本集以及一系列罕见变体。在定量结合试验中，相对于单一标准胎盘ALP（1型）测定每种抗体与每种胎盘ALP的反应性（结合率）。其中六种抗体的研究结果已经报道。本文展示了另外12种抗体的结果，并对这18种抗体的全部系列综合数据进行了分析和讨论。18种抗体中有六种对三种常见等位基因的一种或另一种产物的结合显著降低。在三种情况下，这种区分表现为基本上“全或无”的结合反应。在另外三种情况下，结合差异不太明显，但可以通过结合率的定量比较来证明。定量结合率比较还能够在每种情况下区分杂合子和纯合子。一些抗体与某些罕见变体ALP电泳表型的结合减少。据估计，这一未经筛选的18种抗体系列至少针对胎盘ALP分子表面的九个不同抗原决定簇。结果说明了单克隆抗体区分酶等位基因变体的能力。
