【摘要】 为了增强胆固醇酯转移蛋白(CETP)羧基末端的26肽B细胞抗原表位的免疫原性,以大肠杆菌L-门冬酰胺酶II(AnsB)为载体,通过引入2个强的辅助T细胞表位,构建了融合基因AnsB-TTP-PADRE-CET-PC,并在大肠杆菌BL-21(DE3)表达,经过硫酸铵沉淀、DEAE 52纤维素阴离子交换层析、分子筛凝胶过滤层析,最终得到单一电泳条带纯度的目的蛋白。纯化的融合多肽疫苗免疫新西兰大白兔可诱导产生高水平持续存在的抗CETP抗体,从而抑制了CETP酶活力,显著抑制了兔主动脉斑粥样硬化斑的形成和发展。该融合蛋白有希望作为抗动脉粥样硬化疫苗进一步研究开发。更多还原

【Abstract】 In order to increase the carboxy-terminal B cell epitope immungenicity of human CETP,a fusion protein AnsB-TTP-PADRE-CETPC was successfully constructed,expressed and purified to immunize New Zealand white rabbits for inducing high titers of anti-CETP antibodies to inhibit the formation and development of aortic atheromatous patch.The protein was expressed as soluble protein in Escherichia coli and purified with anion exchange column and by gel filtration chromatography after ammonium sulfate precipitation.After immunizing rabbits with the purified protein,high titer anti-CETP antibodies were induced and it lasted more than nineteen weeks in vivo;patch areas in aorta of rabbits were significantly reduced compared with control rabbits.Therefore,the fusion protein AnsB-TTP-PADRE-CETPC might be further developed into a vaccine against atherosclerosis.更多还原