CAJ | 학술논문

目的比较真核表达、活病毒感染、酵母展示系统用于流感病毒神经氨酸酶(NA) N1亚型抗体筛选的差别,寻找合适的抗体筛选系统.方法将pVRC-VN NA-HAtag质粒转染入293T,通过酵母展示获得重组NA.通过反向遗传学获得重组病毒A/Sichuan/1/2009.用荧光发光法检测NA性能,用流式细胞仪检测NA与5种候选抗体结合情况.结果真核表达、酵母展示系统均可以使NA在表面展示,且具有与活病毒NA相似的特性.在候选的5种抗体中,真核表达以及病毒的NA可与抗体1、5结合,病毒的NA还能与抗体4结合；而酵母展示系统的NA与5种抗体均不能结合.结论真核表达、真病毒感染系统适合NA抗体的筛选,酵母展示系统表达的NA其表面抗体识别的表位有差异.
목적 비교진핵표체、활병독감염、효모전시계통용우류감병독신경안산매(NA) N1아형항체사선적차별,심조합괄적항체사선계통.방법 장pVRC-VN NA-HAtag질립전염입293T,통과효모전시획득중조NA.통과반향유전학획득중조병독A/Sichuan/1/2009.용형광발광법검측NA성능,용류식세포의검측NA여5충후선항체결합정황.결과 진핵표체、효모전시계통균가이사NA재표면전시,차구유여활병독NA상사적특성.재후선적5충항체중,진핵표체이급병독적NA가여항체1、5결합,병독적NA환능여항체4결합；이효모전시계통적NA여5충항체균불능결합.결론 진핵표체、진병독감염계통괄합NA항체적사선,효모전시계통표체적NA기표면항체식별적표위유차이.
Objective To understand if the Neuraminidase (N1) of Influenza A virus at the surface of yeast-displaying system,eukaryotic expression system and the infected cells could be used for anti-NA Abs screening,their activities and bindings to five candidate Abs were assayed.Methods The surface NA expression was obtained by transfecting by recombinant NA constructors with specific tag-labels or live virus infection.The functional activity was measured by the fluorescent assay.Their bindings to the Abs were detected by flow cytometry.Results The surface NAs presenting on the yeast-displaying system and eukaryotic expression system exhibited functional NA activities as the NA at the surface of virus-infected cells which showed affinities to Ab1,4,and 5.The same bindings to Ab1 and 5 were found in the surface NA expressed by eukaryotic expression system while minor binding was observed in the yeast displayed-NA.Conclusion The epitopes of yeast-displayed NA may be different from the NAs present at eukaryotic expression system and the infected cells which more likely suitable for the screening of anti-NA Abs.