抗HIV-2gp36基因工程抗原单克隆抗体的制备、鉴定和初步应用

发布时间：2017-12-28 16:18

本文关键词：抗HIV-2gp36基因工程抗原单克隆抗体的制备、鉴定和初步应用　出处：《河北医科大学》2006年硕士论文　论文类型：学位论文

【摘要】： 目的:由人类免疫缺陷病毒(HIV-1或HIV-2)感染所致的艾滋病,是当今严重威胁人类生命健康的重大疾病,传播广泛迅速,病死率极高,至今尚无治愈方法;检测病毒抗原早期发现感染者,益于对之早期治疗和严格监控,是遏制艾滋病发病和传播的有效手段。我国早期传入和流行的艾滋病毒是HIV-1,针对其代表性病毒抗原p24制备单克隆抗体(简称单抗)的研究已有报道;而近年发现我国也有HIV-2感染和传播,但尚无针对其代表性病毒抗原gp36制备单抗的报道。本研究的目的是,制备稳定产抗gp36基因工程抗原单克隆抗体的杂交瘤细胞株及相应单抗,鉴定细胞株的生物学特性和单抗的免疫学特性及理化特性;初步探索用所制单抗构建检测gp36 ELISA法的适用性;为早期发现HIV-2病毒感染者,提供可资应用的单抗和技术支撑。方法:用基因重组gp36抗原免疫BALB/c小鼠5次,第一次与第二次免疫间隔4周,其后每次免疫间隔2周,融合前三天gp36抗原尾静脉注射进行攻击。取免疫脾细胞与骨髓瘤细胞株Sp2/0,用50%聚乙二醇(PEG)融合,HAT培养基选择培养出融合细胞。用gp36作为抗原包板,间接ELISA法,初筛分泌抗gp36抗体的融合细胞,有限稀释法克隆化培养获得稳定分泌抗gp36单抗的阳性杂交瘤细胞株。以体外连续传30代、液氮中冻存3个月后复苏,测定
[Abstract]:Objective: by the human immunodeficiency virus (HIV-1 or HIV-2) caused by infection of AIDS is threatening the life and health of human beings, spread rapidly, the mortality rate is high, there is no cure for virus antigen detection; early detection of infection, to benefit from early treatment and strict monitoring is an effective means to curb the incidence and the spread of aids. Early introduction and prevailing in China HIV is HIV-1, the viral antigen p24 monoclonal antibody (McAb) research has been reported; and in recent years was found to have the infection and spread of HIV-2 in China, but there is no against its representative antigen gp36 monoclonal antibody reports. The purpose of this study is to prepare stable production of anti gp36 gene engineering antigen monoclonal antibody hybridoma cell lines and corresponding monoclonal antibody, immunological characteristics and physicochemical properties of biological characteristics and identification of monoclonal antibody cell line; preliminary exploration with the adaptability of the McAb in detection of gp36 ELISA construction method for the early detection of HIV-2 virus infection; who provides technical support for the application of monoclonal antibody and.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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