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目的:制备能与鸭乙型肝炎病毒核心抗原(DHB-cAg)特异性结合的单克隆抗体(mAb),使之能特异性地对鸭乙型肝炎病毒(DHBV)进行检测。方法:以原核表达的截短型鸭乙型肝炎病毒核心抗原(DHBcAg1-214)免疫BALB/c小鼠后常规杂交瘤技术进行细胞融合,有限稀释法克隆化,间接酶联免疫吸附试验(ELISA)和免疫组织化学(IHC)筛选、鉴定。结果:筛选出3株(3H4、4A11、5A12)能有效分泌抗鸭乙肝病毒核心抗原的杂交瘤细胞株。该mAb可用于ELISA、IHC和Westernblot研究。结论:获得了3株有效分泌抗核心抗原的mAb,可用于鸭乙肝病毒相关肝病组织与血清的检测。
Objective: To prepare a monoclonal antibody (mAb) capable of specifically binding to duck hepatitis B virus core antigen (DHB-cAg), which can specifically detect duck hepatitis B virus (DHBV). Methods: BALB / c mice were immunized with truncated duck hepatitis B virus core antigen (DHBcAg1-214) and expressed in E.coli BL21 (DE3). The cells were fused by conventional hybridoma technique and cloned by limiting dilution. The indirect enzyme linked immunosorbent assay ) And immunohistochemistry (IHC) screening, identification. Results: Three strains (3H4, 4A11, 5A12) of hybridoma cell lines that can effectively secrete anti-Hepatitis B virus core antigen were screened. The mAb can be used for ELISA, IHC and Western blot studies. Conclusion: Three mAbs that can effectively secrete anti-core antigen were obtained and could be used for the detection of duck hepatitis B virus-associated liver disease and serum.