本文關(guān)鍵詞：抗HIV-2gp36基因工程抗原單克隆抗體的制備、鑒定和初步應(yīng)用　出處：《河北醫(yī)科大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： HIV-2 gp36 雜交瘤 單克隆抗體 ELISA gp36檢測(cè)

【摘要】： 目的:由人類免疫缺陷病毒(HIV-1或HIV-2)感染所致的艾滋病,是當(dāng)今嚴(yán)重威脅人類生命健康的重大疾病,傳播廣泛迅速,病死率極高,至今尚無(wú)治愈方法;檢測(cè)病毒抗原早期發(fā)現(xiàn)感染者,益于對(duì)之早期治療和嚴(yán)格監(jiān)控,是遏制艾滋病發(fā)病和傳播的有效手段。我國(guó)早期傳入和流行的艾滋病毒是HIV-1,針對(duì)其代表性病毒抗原p24制備單克隆抗體(簡(jiǎn)稱單抗)的研究已有報(bào)道;而近年發(fā)現(xiàn)我國(guó)也有HIV-2感染和傳播,但尚無(wú)針對(duì)其代表性病毒抗原gp36制備單抗的報(bào)道。本研究的目的是,制備穩(wěn)定產(chǎn)抗gp36基因工程抗原單克隆抗體的雜交瘤細(xì)胞株及相應(yīng)單抗,鑒定細(xì)胞株的生物學(xué)特性和單抗的免疫學(xué)特性及理化特性;初步探索用所制單抗構(gòu)建檢測(cè)gp36 ELISA法的適用性;為早期發(fā)現(xiàn)HIV-2病毒感染者,提供可資應(yīng)用的單抗和技術(shù)支撐。 方法:用基因重組gp36抗原免疫BALB/c小鼠5次,第一次與第二次免疫間隔4周,其后每次免疫間隔2周,融合前三天gp36抗原尾靜脈注射進(jìn)行攻擊。取免疫脾細(xì)胞與骨髓瘤細(xì)胞株Sp2/0,用50%聚乙二醇(PEG)融合,HAT培養(yǎng)基選擇培養(yǎng)出融合細(xì)胞。用gp36作為抗原包板,間接ELISA法,初篩分泌抗gp36抗體的融合細(xì)胞,有限稀釋法克隆化培養(yǎng)獲得穩(wěn)定分泌抗gp36單抗的陽(yáng)性雜交瘤細(xì)胞株。以體外連續(xù)傳30代、液氮中凍存3個(gè)月后復(fù)蘇,測(cè)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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

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本文編號(hào)：1346536