铜绿假单胞菌疫苗重组蛋白Vac14的纯化方法

Method for purifying pseudomonas aeruginosa vaccine recombinant protein Vac 14

Abstract

The invention discloses a method for purifying pseudomonas aeruginosa (PA) recombinant subunit genetic engineering protein vaccine Vac 14. The Vac 14 protein is obtained in the mode that active functional fragments of pseudomonas aeruginosa antigen molecules are recombined and confused, and escherichia coli genetically engineered bacterium expression is conducted. Technologies including high pressure bacterium breaking, GST affinity chromatography, PP enzyme restriction enzyme digestion, SPHP chromatography, G25 chromatography, Q HP chromatography and the like are adopted for genetic engineering bacteria, and the high-purity pseudomonas aeruginosa (PA) recombinant subunit genetic engineering protein vaccine Vac 14 is obtained. The method is simple and fast in purifying process, easy to amplify and good in repeatability, the obtained target protein is high in purity, and it is proved by animal tests that a mechanism can be effectively stimulated to generate high humoral immune response and a good immune protective effect.
本发明公开了一种铜绿假单胞菌(PA)重组亚单位基因工程蛋白疫苗Vac14的纯化方法。Vac14蛋白为铜绿假单胞菌抗原分子的活性功能片段重组融合,通过大肠杆菌基因工程菌表达获得。采用对基因工程菌进行高压破菌、GST亲和层析、PP酶酶切、SP?HP层析、G?25层析、Q?HP层析等技术,获得高纯度的铜绿假单胞菌(PA)重组亚单位基因工程蛋白Vac14。该发明纯化工艺简捷、容易放大、重复性好,所获目标蛋白纯度高,动物试验证明可有效刺激机体产生较高的体液免疫应答和良好的免疫保护作用。

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