中文摘要：

目的制备抗鸭乙肝病毒core蛋白的单克隆抗体并进行鉴定。方法PCR扩增鸭乙肝病毒（DHBV）core基因片段，构建表达载体pET28a（＋）／DHBVcore。转化诱导表达融合蛋白，用Ni”亲和柱纯化目的蛋白。免疫Balb／c小鼠，取脾细胞与SP2／0骨髓瘤细胞融合，ELISA筛选，采用有限稀释法筛选单克隆杂交瘤细胞并进行细胞克隆。体内诱生法大量制备单克隆抗体，进行抗体的特异性、效价和亚型的鉴定。结果成功构建表达载体pET28a（＋）／DHBVcore，并表达DHBVcore蛋白。获得2株稳定分泌抗DHBVcore抗体的杂交瘤细胞株，分别为2D7和5G10，细胞培养液抗体效价为1：400和1：800。选择5G10细胞株制备单克隆抗体，小鼠腹水抗体效价可达1：320000。鸭肝组织免疫组化结果显示，DHBV病毒载量〉10”copies／mg的肝组织中DHBVcore蛋白的表达明显高于病毒载量〈10。copies／mg的肝组织，而未感染DHBV的鸭肝细胞内不表达DHBVcore蛋白。亚型鉴定结果为IgG2ax链。结论制备并获得了抗DHBVcore蛋白的单克隆抗体，为DHBVcore蛋白的功能研究、诊断试剂的研制与抗病毒治疗研究奠定了基础。

英文摘要：

Objective To prepare and identify monoclonal antibodies against duck hepatitis B virus （DHBV） core protein. Methods DHBV core gene was amplified by PCR to construct a recombinant plasmid pET28a（T）/ DHBV core. The recombinant plasmid was expressed in E. coli BL21 induced by IPTG and DHBV core protein purified by Ni2＋ affinity chromatography was used to immunize 8-week-old female Balb/c Mus musculson. Then after fusion between spleen cells stimulated and SP2/0 myeloma cell, the hybridoma clone against DHBV core protein was obtained by limiting-dilution method and ELISA assay. A large number of antibodies were induced in vivo. The specificity of the antibodies was identified by ELISA assay and immunohistochemistry. Results Recombinant plasmid pET28a（＋）/DHBV core was expressed successfully in E. coil BL21. Two hybridoma clones secreting antibody against DHBV core protein, named 2D7 and 5G10, were obtained. A novel murine monoclonal antibody （5G10） was harvested and it was specific to DHBV core protein. Isotype analysis showed that MAb 5G10 belonged to IgG2a subclass with ~ light chain. Conclusion Monoclonal antibodies against DHBV core protein have been prepared and obtained in this study, which lays the foundation for research on the function of DHBV core protein, development of diagnostic reagents and anti-viral treatment.