CAJ | 학술논문

以Hexa-His和Flag为标签,利用酵母表面展示技术将HIV-1囊膜蛋白gp41展示于酿酒酵母(Saccharomycws cerevisiase)细胞MT8-1表面.流式细胞测定结果表明His标签成功展示于重组酵母细胞MT8-1/pICAS-His和MT8-1/pICAS-His-gp41表面;活性的gp41成功展于重组酵母MT8-1/pICAS-His-gp41表面,荧光显微镜观测结果表明了这一点;Flag标签可以在TM8-1/pICAS-Flag表面检测到,但Flag和gp41均不能在MT8-1/pICAS-Flag-gp41表面检测到活性.利用展示在酵母表面的gp41蛋白建立酵母细胞CbELISA体系,可成功用于检测单克隆抗体,其浓度达到了10 ng/mL,此体系可用于检测血清中的抗体.
이Hexa-His화Flag위표첨,이용효모표면전시기술장HIV-1낭막단백gp41전시우양주효모(Saccharomycws cerevisiase)세포MT8-1표면.류식세포측정결과표명His표첨성공전시우중조효모세포MT8-1/pICAS-His화MT8-1/pICAS-His-gp41표면;활성적gp41성공전우중조효모MT8-1/pICAS-His-gp41표면,형광현미경관측결과표명료저일점;Flag표첨가이재TM8-1/pICAS-Flag표면검측도,단Flag화gp41균불능재MT8-1/pICAS-Flag-gp41표면검측도활성.이용전시재효모표면적gp41단백건립효모세포CbELISA체계,가성공용우검측단극륭항체,기농도체도료10 ng/mL,차체계가용우검측혈청중적항체.
The envelope glycoprotein gp41 of HIV-1 has been successfully displayed on the cell surface of Saccharomyces cerevisiase MT8-1 by cell-surface engineering using Hexa-His and Flag tags for detection, respectively. Flow cytograms showed that His tag was displayed on the surface of MT8-1/pICAS-His and MT8-1/pICAS-His-gp41, the active gp41 could be detected on the cell surface of MT8-1/pICAS-His-gp41 and was confirmed by fluorescence microscopy;Flag tag was detected on the surface of MT8-1/pICAS-Flag, but Flag tag and gp41 were not detected on the surface of MT8-1/pICAS-Flag-gp41 separately. Further, a yeast CbELISA was developed by using the yeast cells displaying gp41 on the cell surface. It was able to detect the monoclonal antibody at the concentration of 10 ng/mL , and also capable of detecting the antibody in the sera.