EB病毒潜伏膜蛋白2A抗原表位的融合表达及其意义

The fusion expression of EBV latent membrane protein-2A antigen epitopes and its significance

为了探讨含有EB病毒潜伏膜蛋白2A(LMP2A)抗原表位片断表达的融合蛋白在鼻咽癌血清学检测中的应用意义,通过重叠延伸PCR方法,合成了3对相互重叠的寡核苷酸引物,涵盖LMP2A的主要的4个抗原表位,将它们拼接在一起构建一个多肽融合基因,克隆到PGEX-4T-2载体中表达融合蛋白,以GST亲和层析柱法纯化融合蛋白,鉴定后以此为抗原检测鼻咽癌患者的血清。结果表明,获得了含EB病毒LMP2A主要的4个抗原表位的融合蛋白(EC2A),蛋白纯度达90%以上,ELISA结果显示鼻咽癌患者血清的检出率为77.9%,正常人群血清为阴性,与常规的VCA-IgA法进行比较,有9份(13.3%)血清VCA-IgA为阴性而EC2A-IgG检出阳性,为鼻咽癌的临床检测提供了新思路,也为后续的单克隆抗体制备奠定了基础。

To explore the significance of fusion proteins of EBV latent membrane protein-2A (LMP2A) antigen in the serological diagnosis for nasopharyngeal carcinoma(NPC),3 pairs of oligonucleotide primers were designed according to the genomic sequences of 4 main antigenic epitopes, the fusion gene containing 4 main LMP2A epitopes was synthesized with overlapping extension PCR and then was cloned into vector pGEX-4T2 for fusion protein to express in E.coli. The recombinant protein was purified by GST chromatography c...