| 不可分型流感嗜血杆菌外膜蛋白P6的纯化及其免疫原性 |
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| 引用本文： | 陈敬,王建华. 不可分型流感嗜血杆菌外膜蛋白P6的纯化及其免疫原性[J]. 中国生物制品学杂志, 2010, 23(5) |
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| 作者姓名： | 陈敬 王建华 |
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| 作者单位： | 北京生物制品研究所细菌研究室,100024 |
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| 摘 要： | 目的纯化不可分型流感嗜血杆菌(NTHi)外膜蛋白P6,并检测其免疫原性。方法常规培养NTHi,根据外膜蛋白P6的特性纯化P6蛋白。Lowry法测定纯化P6蛋白的含量;SDS-PAGE检测纯化P6蛋白的相对分子质量及纯度;IEF-PAGE分析纯化P6蛋白的等电点;琼脂糖双向免疫扩散试验鉴定纯化P6蛋白;以纯化的P6蛋白免疫NIH小鼠,ELISA法检测小鼠血清中IgG抗体水平,并进行小鼠保护性试验。结果纯化的P6蛋白含量为208μg/ml;相对分子质量为16600,纯度为100%;等电点为5.49;琼脂糖双向免疫扩散试验证实所提取的蛋白为P6蛋白;ELISA检测显示,P6蛋白具有良好的免疫原性,免疫小鼠后能够诱导产生针对P6蛋白的抗体,其抗体水平呈剂量依赖关系,最佳免疫剂量为10μg/只,最佳免疫针次为3针;对20LD50NTHiSSIp/1225株攻击的小鼠保护率为80%。结论已纯化了不可分型流感嗜血杆菌外膜蛋白P6,其具有良好的免疫原性和保护效果。
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| 关 键 词： | 嗜血杆菌,流感 细菌外膜蛋白质类 免疫 |
Purification and Immunogenicity of Outer Membrane Protein P6 from Nontypeable Haemophilus influenzae |
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| CHEN Jing,WANG Jian-hua. Purification and Immunogenicity of Outer Membrane Protein P6 from Nontypeable Haemophilus influenzae[J]. Chinese Journal of Bilogicals, 2010, 23(5) |
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| Authors: | CHEN Jing WANG Jian-hua |
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| Affiliation: | CHEN Jing,WANG Jian-hua(National Vaccine , Serum Institute,Beijing 100024,China) |
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| Abstract: | Objective To purify outer membrane protein P6 from nontypeable Haemophilus influenzae(NTHi)and determine its immunogenicity.Methods NTHi was cultured by routine method,from which outer membrane protein P6 was purified according to its characteristics and determined for content by Lowry method,for relative molecular mass and purity by SDS-PAGE,for isoelectric point by IEF-PAGE,and identified by double immunodiffusion test in agarose.NIH mice were immunized with purified P6 protein and determined for serum Ig... |
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| Keywords: | Haemophilus influenzae(NTHi) Bacterial outer membrane protein Immunization |
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