CAJ | 학술논문

目的通过构建免疫旋转生物传感器,建立一种高灵敏度的HIV-1 P24定量检测方法.方法将生物素标记的HIV-1 P24单克隆抗体通过亲和素-生物素-β亚基抗体(Streptavidin-biotin-β antibody)的方式连接到光合作用复合体(Chromatophores)的β亚基上,同时用pH敏感的荧光索F1300标记到Chromatophores上,构建了用于检测P24的免疫旋转生物传感器,并用原核表达的P24抗原对该传感器进行了灵敏度、特异性实验.结果构建的免疫旋转生物传感器可以将P24抗原结合ATPase造成的旋转速率变化转变为光强度的变化,可以高灵敏的检测HIV-1P24抗原,最低检出度可达0.1 fg/ml.结论成功建立了基于免疫生物传感器(Immuno-rotary Biosensor,IRB)系统的HIV-1 P24检测方法,该方法操作简单,灵敏度高,为建立基于分子马达HIV-1 P24定量检测方法打下基础.
목적 통과구건면역선전생물전감기,건립일충고령민도적HIV-1 P24정량검측방법.방법 장생물소표기적HIV-1 P24단극륭항체통과친화소-생물소-β아기항체(Streptavidin-biotin-β antibody)적방식련접도광합작용복합체(Chromatophores)적β아기상,동시용pH민감적형광색F1300표기도Chromatophores상,구건료용우검측P24적면역선전생물전감기,병용원핵표체적P24항원대해전감기진행료령민도、특이성실험.결과 구건적면역선전생물전감기가이장P24항원결합ATPase조성적선전속솔변화전변위광강도적변화,가이고령민적검측HIV-1P24항원,최저검출도가체0.1 fg/ml.결론 성공건립료기우면역생물전감기(Immuno-rotary Biosensor,IRB)계통적HIV-1 P24검측방법,해방법조작간단,령민도고,위건립기우분자마체HIV-1 P24정량검측방법타하기출.
Objective To build a high sensitive method to detect HIV-1 P24 antigen quantitatively by Immuno-rotary Biosensor (IRB) based on F0F1-ATPase. Methods The immuno-rotary biosensor based on F0F1-ATPase in chromatophores for detecting HIV-1 P24 was assembled by label F0F1-ATPase chromatophores with the biotin-labled HIV-1 P24 antibody through the streptavidin-biotin-β antibody and the pH sensitivity of fluorescence F1300. Then it's sensitivity and specificity was tested by the prokaryotic expressed HIV-1 P24 antigen. Results Immuno-rotary biosensor based on F0F1-ATPase in chromatophores can detect P24 antigen sensitivily, the lowest level that could be detected was 0.1 fg/ml. Conclusion IRBbased system was successfully assembled and used for the detection of P24 antigen, the rapid and sensitive technique will be useful for detecting HIV-1 P24 antigen quantitatively.