中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2012年
9期
838-842
,共5页
张晓丽%韩晓旭%代娣%包名家%张子宁%赵敏%年华%尚红
張曉麗%韓曉旭%代娣%包名傢%張子寧%趙敏%年華%尚紅
장효려%한효욱%대제%포명가%장자저%조민%년화%상홍
HIV-1%表位%肽库%HIV感染%中和抗体
HIV-1%錶位%肽庫%HIV感染%中和抗體
HIV-1%표위%태고%HIV감염%중화항체
HIV-1%Epitopes%Peptide library%HIV infections%Antibodies,neutralizing
目的 以含有高水平HIV-1中和抗体的血浆为配体筛选HIV-1中和抗体模拟性抗原表位.方法 为获取中和抗体识别的模拟表位,采用多轮感染外周血单个核细胞( PBMC)中和实验检测11例HIV感染缓慢进展者血浆中和抗体水平,利用噬菌体随机肽库对具有高水平中和抗体的血浆进行生物淘筛,ELISA方法鉴定阳性克隆.阳性克隆测序后与HIV Env gp41序列进行了同源性分析,应用Local Blast软件分析中和抗体模拟表位.结果 3轮淘筛之后,得到22个阳性克隆.分析发现包括C8等12个可能的中和抗体模拟表位,位于gp41Ⅱ类功能区的C40对HIV-1血浆具有较高的抑制率.结论 利用缓慢进展者血浆IgG抗体筛选噬菌体随机肽库,能够有效的获得多个HIV-1中和抗体相关抗原模拟表位.
目的 以含有高水平HIV-1中和抗體的血漿為配體篩選HIV-1中和抗體模擬性抗原錶位.方法 為穫取中和抗體識彆的模擬錶位,採用多輪感染外週血單箇覈細胞( PBMC)中和實驗檢測11例HIV感染緩慢進展者血漿中和抗體水平,利用噬菌體隨機肽庫對具有高水平中和抗體的血漿進行生物淘篩,ELISA方法鑒定暘性剋隆.暘性剋隆測序後與HIV Env gp41序列進行瞭同源性分析,應用Local Blast軟件分析中和抗體模擬錶位.結果 3輪淘篩之後,得到22箇暘性剋隆.分析髮現包括C8等12箇可能的中和抗體模擬錶位,位于gp41Ⅱ類功能區的C40對HIV-1血漿具有較高的抑製率.結論 利用緩慢進展者血漿IgG抗體篩選噬菌體隨機肽庫,能夠有效的穫得多箇HIV-1中和抗體相關抗原模擬錶位.
목적 이함유고수평HIV-1중화항체적혈장위배체사선HIV-1중화항체모의성항원표위.방법 위획취중화항체식별적모의표위,채용다륜감염외주혈단개핵세포( PBMC)중화실험검측11례HIV감염완만진전자혈장중화항체수평,이용서균체수궤태고대구유고수평중화항체적혈장진행생물도사,ELISA방법감정양성극륭.양성극륭측서후여HIV Env gp41서렬진행료동원성분석,응용Local Blast연건분석중화항체모의표위.결과 3륜도사지후,득도22개양성극륭.분석발현포괄C8등12개가능적중화항체모의표위,위우gp41Ⅱ류공능구적C40대HIV-1혈장구유교고적억제솔.결론 이용완만진전자혈장IgG항체사선서균체수궤태고,능구유효적획득다개HIV-1중화항체상관항원모의표위.
Objective To screen mimetic HIV-1 neutralizing epitopes from plasma with high level neutralizing antibody,and to provide useful information for further study of the interaction between antigen and antibody.Methods In order to gain neutralizing antibody recognized mimotopes, we detected neutralizing antibodies levels of 11 HIV-1 infected slow progressors by PBMC-based neutralization assays.High-titer HIV-neutralizing antibodies from plasma of SPs was used as the ligand for biopanning by phage-displayed random peptide library.Positive phage clones was evaluated by ELISA,sequenced,and analyzed for homology to HIV-1 env by local BLAST to deduce the neutralizing peptide.Results Twenty-two clones were obtained consistent with requirement through three rounds biopanning.After comparison analysis,twelve clones include C8 were obtained as mimotopes of neutralizing antibody,C40 located in gp41 Ⅱ cluster with the highest titer by inhibition ratio may be as neutralizing epitope.Conclusion By the use of IgG antibodies from SPs to screen the phage random polypeptide library,one can acquire multiple phage mimetic peptides of HIV related antigen epitope.(Chin J Lab Med,2012,35:838-842 )
