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目的：研究β趋化因子受体５（ＣＣＲ５）ＮＨ２端膜外第一襻结构域的功能及其特异抗体Ｆ（ａｂ′）２的制备。方法：计算机分析趋化因子超基因家族，定位超基因家族中同源顺序最低的结构域ＮＨ２端膜外结构域，ＰＣＲ扩增出该结构域１１４个核苷酸序列。构建融合表达载体ｐＧＥＸ－ＩＮ／ＮＲ５，经测序鉴定正确后，在Ｅ．ｃｏｌｉ中表达，经纯化后免疫新西兰兔。蛋白Ａ亲和层析和胰蛋白酶消化ＩｇＧ，经Ｓｅｐｈａｒｏｓｅ－１２柱层析制备Ｆ（ａｂ′）２。结果：经还原和非还原聚丙烯酰胺凝胶电泳和ＦＡＸ分析证明得到抗ＣＣＲ５ＮＨ２端的特异性抗体。结论：本方法为一简捷快速的特定功能结构域抗体Ｆ（ａｂ′）２制备方法，对研究该基因在体内的表达及生物学功能提供了重要的实验材料，同时也对超基因家族中亚家族特异抗体研制提供了一种研究思路和方法。
AIM: To investigate the function of the first 外 domain outside the membrane of β-chemokine receptor 5 (CCR5) NH2 and the preparation of its specific antibody F (ab ’) 2. Methods: The chemotactic supergene gene family was analyzed by computer. The NH2-terminal extracellular domain of the domain with the lowest homologous sequence in the supergene family was located and 114 nucleotides of this domain were amplified by PCR. The fusion expression vector pGEX-IN / NR5 was constructed and verified by sequencing. expressed in E. coli, purified and immunized New Zealand rabbits. Protein A affinity chromatography and tryptic digestion of IgG, F (ab ’) 2 was prepared by Sepharose-12 column chromatography. Results: Reduced and non-reduced polyacrylamide gel electrophoresis and FAX analysis demonstrated that specific antibodies against the CCR5NH2 end were obtained. Conclusion: This method is a simple and rapid method for the preparation of specific functional domain antibody F (ab ’) 2. It provides an important experimental material for studying the expression and biological function of this gene in vivo. In addition, Family-specific antibody provides a research ideas and methods.