| HBV颗粒的纯化及生物学活性鉴定 |
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| 引用本文： | 曾兴光,石统东,韩俊峰,邹丽云,谢谆怡,刘宏利,吴玉章. HBV颗粒的纯化及生物学活性鉴定[J]. 免疫学杂志, 2010, 0(4) |
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| 作者姓名： | 曾兴光 石统东 韩俊峰 邹丽云 谢谆怡 刘宏利 吴玉章 |
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| 作者单位： | 第三军医大学.基础医学部全军免疫学研究所;重庆医科大学附属第二医院病毒性肝炎研究所; |
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| 摘 要： | 目的优化纯化HBV患者血清中HBV颗粒的技术方法。方法在蔗糖介质中,分别以100 000 g和70 000 g进行不连续蔗糖梯度(60%,45%,35%,25%,15%)离心5 h,离心后,分段吸取6份收集液,PCR荧光定量检测各组分HBV滴度;收集前5份收集液进行超滤,去蔗糖,PCR荧光定量检测HBV超滤液;电镜观察70 000g纯化后的HBV超微形态;70000 g纯化后的HBV颗粒感染树鼩肝细胞,检验70 000 g纯化后的HBV感染活性。结果100 000 g离心造成大量HBV在离心过程中损失,收获率只有21%,70 000g离心不仅降低了离心中的病毒损失,而且可有效对HBV和血清进行分离,收获率达到78%以上,较100 000g还提高3.7倍多。电镜可观察到纯化后病毒液中有大量的病毒颗粒;纯化后的HBV颗粒感染活性良好,可有效感染原代树鼩肝细胞。结论70 000 g不连续蔗糖梯度离心可有效分离纯化HBV病毒,较100 000 g收获率明显提高,纯化后的HBV不但具有典型的HBV特征,还具有良好的生物学感染活性。
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| 关 键 词： | 乙型肝炎病毒 不连续蔗糖梯度离心 纯化 树鼩肝细胞 |
Purification of HBV virions and validation of its biological activity |
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| ZENG Xingguang,SHI Tong olong,HAN Junfeng,ZOU Liyun,XIE Zunyi,LIU Lihong,Wu yuzhang Institute of Immunology,Third Military Medical University,Chongqing ,China. Purification of HBV virions and validation of its biological activity[J]. Immunological Journal, 2010, 0(4) |
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| Authors: | ZENG Xingguang SHI Tong olong HAN Junfeng ZOU Liyun XIE Zunyi LIU Lihong Wu yuzhang Institute of Immunology Third Military Medical University Chongqing China |
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| Affiliation: | ZENG Xingguang,SHI Tong olong,HAN Junfeng,ZOU Liyun,XIE Zunyi,LIU Lihong,Wu yuzhang Institute of Immunology,Third Military Medical University,Chongqing 40038,China |
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| Abstract: | In this study,we aimed to optimize the method for purifying HBV virions from serum of HBV patients.HBV virions were purified by using discontinuous sucrose density gradient(60%,45%,35%,25%,15%sucrose solutions) centrifugation at 100 000 g and 70 000 g for 5 h.After the centrifugation,six fractions were collected and real time fluorescent quantitative polymerase chain reaction was used to characterize the fractions containing HBV virions.The first 5 fractions were mixed and ultrafiltrated to remove sucrose, ... |
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| Keywords: | HBV Discontinuous sucrose density gradient Purification Tupaia hepatocyte |
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