抗猪流行性腹泻病毒噬菌体单链抗体库的构建及筛选

Construction and Screening of a Phage Display scFv Library against Porcine Epidemic Diarrhea Virus

构建抗猪流行性腹泻病毒(PEDV)噬菌体单链抗体库,筛选鉴定抗猪流行性腹泻病毒的单链抗体。用猪流行性腹泻疫苗免疫发病耐过的猪,3次免疫后采血,分离淋巴细胞,提取淋巴细胞的总RNA,分别扩增猪IgG重链和轻链可变区,将其组装成scFv基因,连接到噬菌体质粒。进行3轮吸附-洗脱-富集选淘抗猪流行性腹泻病毒的噬菌体单链抗体库。结果构建了库量为9.5×10~9 pfu/mL抗猪流行性腹泻病毒的单链抗体库,筛选并得到1株亲和活性较好的抗猪流行性腹泻病毒的单链抗体。成功构建了抗猪流行性腹泻病毒的噬菌体单链抗体库,并获得抗猪流行性腹泻病毒的单链抗体,可为猪流行性腹泻单克隆抗体的制备提供参考。

To construct the phage display scFv library against porcine epidemic diarrhea virus(PEDV)and to screen hight specificity antibody for PEDV.Swine were immunized with vaccine,of which the peripheral blood samples were collected after the3rd immunization.Total RNA was isolated from lymphocytes,which was extracted for amplification of heavy chain and light chain of IgG.The obtained scFv gene was inserted into expression vector.After three rounds of binding-elution-enrichment,we obtained a diversity of approximately9.5×109pfu/ml anti-PEDV scFv antibody library and screened a hight specificity scFv antibody for PEDV.The phage antibody library against porcine epidemic diarrhea virus was successfully constructed and provided a reference for the preparation of monoclonal antibody against swine epidemic diarrhea.