论文部分内容阅读
目的制备甲型流感H1N1(2009)流感病毒M2e单克隆抗体,并建立检测M2e的双抗体夹心ELISA方法。方法采用碳化二亚胺法将人工合成的M2e多肽分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)进行偶联后作为免疫原,利用杂交瘤细胞技术制备抗A型流感M2e单克隆抗体并进行抗体的鉴定,建立检测M2E的双抗体夹心ELISA方法。结果制备了3株甲型流感H1N1(2009)流感病毒M2e单克隆抗体,经鉴定单克隆抗体均为IGg1a亚型。利用制备的M2e单克隆抗体建立和优化甲型流感M2e检测双抗体夹心ELISA方法,特异性符合临床要求,灵敏度为19ng/ml。结论制备了甲型流感H1N1(2009)M2e单克隆抗体,建立的M2e检测双抗体夹心ELISA方法具有较高的灵敏度和特异性,为研究M2e单克隆抗体作用机制奠定了基础。
Objective To prepare the M2e monoclonal antibody of influenza A H1N1 (2009) and to establish a sandwich ELISA method for detecting M2e. Methods M2e polypeptides synthesized by carbodiimide method were conjugated with bovine serum albumin (BSA) and ovalbumin (OVA) respectively as immunogens. The monoclonal anti-M2e monoclonal antibody was prepared by hybridoma cell technology Antibodies and antibody identification, the establishment of detection of M2E double antibody sandwich ELISA method. Results Three strains of M2e monoclonal antibodies against influenza A (H1N1) (H1N1) virus were prepared. All monoclonal antibodies were identified as IGg1a subtypes. The established M2e monoclonal antibody was used to establish and optimize the sandwich ELISA for M2e detection of influenza A virus. The sensitivity was 19ng / ml according to the clinical requirement. Conclusions The monoclonal antibody against influenza A H1N1 (2009) M2e was prepared and the established sandwich ELISA method for detecting M2e with high sensitivity and specificity provided a basis for studying the mechanism of M2e monoclonal antibody.