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脑膜炎球菌P64K蛋白作为新型抗原载体的研究进展 被引量:1
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作者 熊向华 刘志敏 《中国生物工程杂志》 CAS CSCD 2003年第10期19-22,共4页
P64K是来自奈瑟氏属脑膜炎球菌的一种外层膜蛋白 ,发现于脑膜炎恢复期患者和接种过古巴VA MENGOC BC疫苗的个体血清内。编码P64K蛋白的基因称lpdA ,其编码序列长 1 782bp。P64K蛋白含 5 93个氨基酸 ,由两个结构域组成 ,有二氢硫辛酸酰... P64K是来自奈瑟氏属脑膜炎球菌的一种外层膜蛋白 ,发现于脑膜炎恢复期患者和接种过古巴VA MENGOC BC疫苗的个体血清内。编码P64K蛋白的基因称lpdA ,其编码序列长 1 782bp。P64K蛋白含 5 93个氨基酸 ,由两个结构域组成 ,有二氢硫辛酸酰胺酰基脱氢酶活性。P64K是抗脑膜炎疫苗的候选者。因为P64K的高分子量和强免疫原性 。 展开更多
关键词 脑膜炎球菌 p64k蛋白 抗原载体 免疫原性
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Construction and evaluation of anti-gastrin immunogen based on P64K protein 被引量:1
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作者 Xiang-Hua Xiong Hong-Liang Zhao Chong Xue Wei Zhang Bing-Fen Yang Xue-Qin Yao Zhi-Min Liu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第13期2040-2046,共7页
AIM: To construct two kinds of anti-gastrin immunogen based on P64K protein from Neisseria rneningitids and to compare their immunogenic effect. METHODS: G17P64K gene was cloned and ligated into pET28a plasmid, then... AIM: To construct two kinds of anti-gastrin immunogen based on P64K protein from Neisseria rneningitids and to compare their immunogenic effect. METHODS: G17P64K gene was cloned and ligated into pET28a plasmid, then transformed into BL21(DE3). After inoculation of LB medium and IPTG induction, the recombinant protein was solubly expressed at a high level. The purification of G17P64K fusion protein was similar to that of P64K. An initial step of purification consisting of 30% saturated ammonium sulfate precipitation was done, Additional fine optimizations included phenyl-sepharose, G200 Sephadex gel filtration and Q-sepharose anion exchanger chromatography. Highly purified protein was obtained and sequenced at the N-terminal amino acid residues. Polypeptide was synthesized by Fmoc solid phase chemical method and cross-linked to carrier protein P64K and DT mutant by MBS method and then the rabbit anti-gastrin 17 antibody was prepared by immunizing rabbit with cross-linked and fused protein. The titer and the activity in vitro of antibody were assessed. RESULTS: G17P64K gene and the recombinant bacteria were obtained. After four steps purification, protein sample that has the purity above 90% was achieved. At the 84^th day after the first immunization, the titer of antibody against cross-linked protein reached 51 200. Evaluation of the antibody in vitro manifested that it had a high inhibitory activity on the growth of tumor cell SW480. CONCLUSION: The P64K-polypeptide cross-linked immunogen immunized rabbit and achieved a higher titer antibody against gastrin 17 than the G17P64K fusion protein immunogen, which could inhibit the growth of the tumor cell SW480. 展开更多
关键词 GASTRIN p64k protein therapeutic vaccine
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