中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2009年
9期
851-853
,共3页
邓郁青%张征峥%王平%丁军颖%张红中%王润田
鄧鬱青%張徵崢%王平%丁軍穎%張紅中%王潤田
산욱청%장정쟁%왕평%정군영%장홍중%왕윤전
HIV%单克隆抗体%ELISA
HIV%單剋隆抗體%ELISA
HIV%단극륭항체%ELISA
HIV%Monoclonal antibody%ELISA
目的 建立多种单抗联合早期检测HIV抗原的夹心ELISA方法.方法 以SAS盐析沉淀法和亲和层析法纯化抗HIV-1 p24、gp41、gp120及抗HIV-2 gp36的腹水型单克隆抗体(McAb),用高碘酸钠法将纯化的McAb以HRP进行标记.建立针对单个抗原的双抗体夹心ELISA法,对其灵敏度及特异性进行检测.将筛选得到的4株捕获McAb按比例混合作为捕获抗体,4株酶标McAb按比例混合作为检测抗体,建立多种单抗联合检测HIV抗原的夹心ELISA方法,检测混合HIV抗原.结果 按确定的最优反应条件建立的多种McAb联合夹心ELISA方法,检测到的最高稀释度的HIV混合抗原中各抗原的终浓度分别为:重组HIV-1 p24:0.625 pg/ml,gp41:6.25 ng/ml,gp120:6.25 ng/ml;HIV-2 gp36:9.25 ng/ml.结论 建立了具有高度敏感性的鸡尾酒式多种单抗联合检测HIV抗原的夹心ELISA法,为早期榆测HIV抗原提供了新的思路,为后续的研究奠定了一定基础.
目的 建立多種單抗聯閤早期檢測HIV抗原的夾心ELISA方法.方法 以SAS鹽析沉澱法和親和層析法純化抗HIV-1 p24、gp41、gp120及抗HIV-2 gp36的腹水型單剋隆抗體(McAb),用高碘痠鈉法將純化的McAb以HRP進行標記.建立針對單箇抗原的雙抗體夾心ELISA法,對其靈敏度及特異性進行檢測.將篩選得到的4株捕穫McAb按比例混閤作為捕穫抗體,4株酶標McAb按比例混閤作為檢測抗體,建立多種單抗聯閤檢測HIV抗原的夾心ELISA方法,檢測混閤HIV抗原.結果 按確定的最優反應條件建立的多種McAb聯閤夾心ELISA方法,檢測到的最高稀釋度的HIV混閤抗原中各抗原的終濃度分彆為:重組HIV-1 p24:0.625 pg/ml,gp41:6.25 ng/ml,gp120:6.25 ng/ml;HIV-2 gp36:9.25 ng/ml.結論 建立瞭具有高度敏感性的鷄尾酒式多種單抗聯閤檢測HIV抗原的夾心ELISA法,為早期榆測HIV抗原提供瞭新的思路,為後續的研究奠定瞭一定基礎.
목적 건립다충단항연합조기검측HIV항원적협심ELISA방법.방법 이SAS염석침정법화친화층석법순화항HIV-1 p24、gp41、gp120급항HIV-2 gp36적복수형단극륭항체(McAb),용고전산납법장순화적McAb이HRP진행표기.건립침대단개항원적쌍항체협심ELISA법,대기령민도급특이성진행검측.장사선득도적4주포획McAb안비례혼합작위포획항체,4주매표McAb안비례혼합작위검측항체,건립다충단항연합검측HIV항원적협심ELISA방법,검측혼합HIV항원.결과 안학정적최우반응조건건립적다충McAb연합협심ELISA방법,검측도적최고희석도적HIV혼합항원중각항원적종농도분별위:중조HIV-1 p24:0.625 pg/ml,gp41:6.25 ng/ml,gp120:6.25 ng/ml;HIV-2 gp36:9.25 ng/ml.결론 건립료구유고도민감성적계미주식다충단항연합검측HIV항원적협심ELISA법,위조기유측HIV항원제공료신적사로,위후속적연구전정료일정기출.
Objective To establish a sandwich ELISA for early detection of HIV antigens using a mixture of monoclonal antibodies (McAb). Methods The ascites McAbs (anti-HIV-1 p24, anti-HIV-1 gp41, anti-HIV-1 gp120 and anti-HIV-2 gp36) were purified by the SAS and the affinity chromatography,and then were labeled with HRP by sodium metaperiodate. The establishing of sandwich ELISA for detecting the single HIV antigen and the tests of specificity and sensitivity of these systems were performed in advance.A proper ratio mixture of four screened McAbs was used as the capture antibody and a proper ratio mixture of four labeled antibodies was used as the detecting antibody. The method of using sandwich ELISA to detect HIV antigens was set up with these McAbs. Results The sensitivity of this method detecting HIV antigens are:0.625 pg/ml HIV-1 p24, 6.25 ng/ml HIV-I gp41,6.25 ng/ml HIV-I gp120 and 9.25 ng/mi HIV-2 gp36 in mixed HIV antigens. Conclusion The method of using several McAbs mixture in sandwich ELISA detecting HIV antigens was established an excellent sensitivity, which provides a novel idea for early detec-ting the HIV antigen.
