一株抗腮腺炎病毒Fab抗体克隆的筛选

SELECTION OF FAB PHAGE ANTIBODIES CLONING AGAINST MUMPS VIRUS

[目的]获得抗腮腺炎病毒Fab段基因工程抗体。[方法]将腮腺炎病毒抗原(MUV-Ag)包被在固相ELISA板中,采用"吸附-洗脱-扩增"的方法,对已构建的抗腮腺炎病毒噬菌体抗体库进行富集。从富集后的抗体库中筛选抗腮腺炎病毒阳性克隆,并交叉筛选鉴定其特异性。[结果]在对176个克隆筛选后,再用麻疹病毒抗原(MEV-Ag)、混合副流感病毒抗原(mPIV-Ag)和呼吸道合胞病毒抗原(RSV-Ag)进行交叉筛选,最终获得特异性抗MUV-Ag阳性克隆2个(B206和D210)。对其中1株B206序列分析表明它的κ轻链V基因属于VκⅠ(O12)亚群,J基因属于Jκ1;γ链V基因属于VH1-69亚群,D基因来自D3-10,J基因则来自JH6。[结论]用腮腺炎病毒抗原成功地从抗体库中筛选到特异性阳性克隆,构建的噬菌体抗体库是有效的。

[Objective]To obtain Fab phage genetically engineered antibodies against mumps virus.[Methods]MUV-Ag was invested in solid phrase of ELISA,and phage antibody library against mumps virus constructed was enriched by using adsorption-elution-amplification method.The positive clones of against mumps virus were selected from the antibody library which had been enriched.The positive clones were cross-selected as well as the specificity of them were detected.[Results]After selecting 176 clones,these clones were cross-selected by using MEV-Ag,mPIV-Ag and RSV-Ag.Finally we obtained two positive clones of specific MUV-Ag（B206 and D210） .The sequence analysis of one strain of B206 showed that the V gene in κ-light chain derived from VκⅠsubgroup as well as the J gene from Jκ1;the V,D and J geneS in γchain were respectively derived from VH1-69 D3-10 and JH6 subgroups.[Conclusion]The specific positive clones were successfully selected from antibody library by using mumps virus antigen,which indicates the phage antibody library constructed is effective.