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三种新型抗丙型肝炎病毒核心蛋白单克隆抗体的特性分析

Characterization of three novel monoclonal antibodies against hepatitis C virus core protein.

作者信息

Moradpour D, Wakita T, Tokushige K, Carlson R I, Krawczynski K, Wands J R

机构信息

Molecular Hepatology Laboratory, Massachusetts General Hospital Cancer Center, Charlestown, MA 02129, USA.

出版信息

J Med Virol. 1996 Mar;48(3):234-41. doi: 10.1002/(SICI)1096-9071(199603)48:3<234::AID-JMV4>3.0.CO;2-9.

Abstract

Three novel monoclonal antibodies (MAbs) were established against a recombinant hepatitis C virus (HCV) core protein derived from cloned genotype 1b HCV cDNA. MAbs C7-50 and C8-59 recognize a conserved linear epitope represented by amino acid residues 21 to 40 of the nucleocapsid protein. MAb C8-48 is directed against a strain-specific conformational epitope located within the first 82 amino acids. A sensitive two-site MAb-based immunoradiometric assay was established using antibodies directed against distinct epitopes on the nucleocapsid protein. Processed 21 kDa core protein was detected by immunoblotting in human hepatocellular carcinoma cell lines and primary adult rat hepatocytes transfected with a cytomegalovirus promoter-driven expression construct. Immunofluorescence microscopy studies revealed a granular and vesicular cytoplasmic staining pattern. MAb C7-50 was used successfully to detect HCV core antigen in chronically infected chimpanzee liver tissue. These MAbs represent important reagents for the study of HCV biology and for the development of immunodiagnostic assays.

摘要

针对源自克隆的1b型丙型肝炎病毒(HCV)cDNA的重组HCV核心蛋白,制备了三种新型单克隆抗体(MAb)。单克隆抗体C7-50和C8-59识别由核衣壳蛋白氨基酸残基21至40代表的保守线性表位。单克隆抗体C8-48针对位于前82个氨基酸内的菌株特异性构象表位。使用针对核衣壳蛋白上不同表位的抗体建立了一种灵敏的基于双位点单克隆抗体的免疫放射分析方法。通过免疫印迹法在用人巨细胞病毒启动子驱动的表达构建体转染的人肝癌细胞系和原代成年大鼠肝细胞中检测到加工后的21 kDa核心蛋白。免疫荧光显微镜研究揭示了颗粒状和小泡状细胞质染色模式。单克隆抗体C7-50成功用于检测慢性感染黑猩猩肝脏组织中的HCV核心抗原。这些单克隆抗体是研究HCV生物学和开发免疫诊断分析方法的重要试剂。

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