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寻常型天疱疮抗原 (pemphigusvulgarisantigen ,PVA )是桥粒中相对分子质量为 130 0 0 0的糖蛋白 ,其分子由细胞外区(extracellulardomain ,EC )、跨膜区和细胞内区三部分组成 ,主要发挥作用的区域在细胞外区。为了了解PVA细胞外区的致病性位点及其相应抗体的致病作用 ,制备抗血清是必需环节。本文在已重组表达的PVAEC1 2融合蛋白的基础上进行蛋白纯化 ,并免疫家兔 ,使之产生针对融合蛋白的特异性抗血清 ,经双向免疫扩散实验 ,其效价为 1:16～ 1:32。采用辛酸沉淀法和DEAE 5 2 纤维素阴离子交换层析进行抗血清IgG抗体成分的提纯 ,总量为 12 5～ 2 2 0mg ,经间接免疫荧光检测 ,其滴度为 1:40。从而获得高滴度、高特异性的兔抗PVAEC1 2融合蛋白抗血清IgG抗体成分。
Pemphigus vulgarisantigen (PVA) is a glycoprotein with a molecular weight of 130,000 in desmosomes. Its molecule consists of extracellular domain (EC), transmembrane region and intracellular region. The main area of action is in the extracellular region. In order to understand the pathogenicity of PVA extracellular region and its corresponding antibody pathogenic role, the preparation of antisera is an essential part. In this paper, the protein was purified on the basis of the recombinant protein of PVAEC1 2 and immunized rabbits to produce specific antiserum against the fusion protein. The titer of the fusion protein was 1: 16-1: 32. The antiserum IgG antibody was purified by caprylic acid precipitation and DEAE 5 2 anion exchange chromatography. The total amount was 125 ~ 220 mg. The titer was 1:40 by indirect immunofluorescence assay. Thereby obtaining a high titer, high specificity rabbit anti-PVAEC1 2 fusion protein anti-serum IgG antibody component.