Selection of a peptide mimicking neutralization epitope of hepatitis E virus with phage peptide disp

来源 :世界胃肠病学杂志(英文版) | 被引量 : 0次 | 上传用户:lialianing
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
AIM: To select the peptide mimicking the neutralization epitope of hepatitis E virus which bound to non-type-specific and conformational monoclonal antibodies (mAbs) 8C11 and 8H3 fromed 7-peptide phage display library, and expressed the peptide recombinant with HBcAg in E.coli, and to observe whether the recombinant HBcAg could still form virus like particle (VLP) and to test the activation of the recombinant polyprotein and chemo-synthesized peptide that was selected by mAb 8H3.METHODS: 8C11 and 8H3 were used to screen for binding peptides through a 7-peptide phage display library. After 4rounds of panning, monoclonal phages were selected and sequenced. The obtained dominant peptide coding sequences was then synthesized and inserted into amino acid 78 to 83 of hepatitis B core antigen (HBcAg), and then expressed in E. coli. Activity of the recombinant proteins was detected by West blotting, VLPs of the recombinant polyproteins were tested by transmission electron microscopy and binding activity of the chemo-synthesized peptide was confirmed by BIAcore biosensor.RESULTS: Twenty-one positive monoclonal phages (10for 8CL1, and 11 for 8H3) were selected and the inserted fragments were sequenced. The DNA sequence coding for the obtained dominant peptides 8C11 (N′-His-Pro-Thr-LeuLeu-Arg-Ile-C′, named 8C11A) and 8H3 (N′-Ser-Ile-LeuPro- Tyr-Pro-Tyr-C′, named 8H3A) were then synthesized and cloned to the HBcAg vector, then expressed in E. coli.The recombinant proteins aggregated into homodimer or polymer on SDS-PAGE, and could bind to mAb 8C11 and 8H3 in West blotting. At the same time, the recombinant polyprotein could form virus like particles (VLPs), which could be visualized on electron micrograph. The dominant peptide 8H3A selected by mAb 8H3 was further chemosynthesized, and its binding to mAb 8H3 could be detected by BIAcore biosensor.CONCLUSION: These results implicate that conformational neutralizing epitope can be partially modeled by a short peptide, which provides a feasible route for subunit vaccine development.
其他文献
期刊
期刊
目的:探讨神经元活化在癫(癎)发生、发展中的作用及托吡酯对其的影响.方法:采用戊四氮制备慢性癫(癎)模型,利用托吡酯干扰,选取不同时间点,观察大鼠行为学变化及神经细胞黏附
目的:观察中西医结合治疗病毒性心肌炎的临床疗效。方法:将60例病毒性心肌炎患者随机分为治疗组和对照组各30例,两组患者均采用常规西医治疗,治疗组患者在常规治疗基础上加服
研究目的:深入探究为老年性白内障患者实施优质护理的应用价值.护理方法:从本院选取60例诊断出老年性白内障的患者作为本次的研究对象,按照平均分配的方式将他们分为两组,每
期刊
期刊
目的:探究中西医结合对于骨质疏松胸腰椎压缩骨折疼痛的治疗效果.方法:选取我院骨质疏松性胸腰椎压缩骨折患者92例,分为对照组和观察组,各46例,两组患者采用不同的治疗方式,
期刊
@@
1 概述rn生物反馈疗法是一种有意识的放松疗法,它借助于传感器把采集到的生理心理活动信息加以处理和放大,及时转换成人们熟悉的视觉和听觉信号,让人们“感觉”到自己内脏器