中文摘要：

为制备能够区分禽白血病A亚群和J亚群病毒的单克隆抗体,将J亚群病毒gp85基因构建到原核表达载体上,并在大肠杆菌BL21中表达携带His标签的禽白血病J亚群GP85融合蛋白,用复性纯化的融合蛋白作为免疫原免疫BALB/c小鼠。4次免疫后取免疫小鼠的脾细胞与骨髓瘤细胞（SP2/0）进行融合,经过3次亚克隆后获得1株稳定分泌针对GP85蛋白的杂交瘤细胞株,命名为386。经间接ELISA测定,小鼠腹水效价为6.4×10^5,亲和力解离常数（kD）为2.18×10^-9,这株单抗的亚型为IgGl。通过Western Blot和IFA实验证实该株单抗是针对J亚群病毒蛋白GP85的特异性抗体。初步确定此株单克隆抗体的抗原识别区位于GP85蛋白N端的1-50位氨基酸。该株单抗的制备为ALV-J病毒抗原检测以及致病机理的研究奠定了基础。

英文摘要：

To develop monoclonal antibodies（McAbs） that can distinguish avian leukemia subgroup A virus from subgroup J virus,we cloned gp85 gene into prokaryotic expression vectors pET-32 a and pGEX-6P-1.The expression vectors were transformed into E.coli BL21 for expression of GP85.The purified and refolded fusion protein GP85-His was used as an immunogen to vaccinate BALB/c mice.Then,we fused myeloma cells SP2/0 with splenocytes from the immunized BALB/c mice after four times of immunization.After three rounds of subcloning,we obtained a hybridoma cell clone that stably secreted McAbs against GP85 and named 3B6.Antibody titers were 6.4×10^5 as examined by indirect ELISA in ascite fluid of mice inoculated intraperitoneally with monoclonal antibody-producing hybridoma cells.The dissociation constant（kD） of the monoclonal antibody was determined to be2.18×10^-9 and the isotype was IgG1.Specific recognition of GP85 by anti-GP85 monoclonal antibody was validated by Western Blot and indirect Fluorescence Antibody assay.The epitope in GP85 that was recognized by 3B6 was located between 1 to 50 aa as demonstrated by Western Blot.The McAb will be helpful to the examination of ALV-J antigen and to elucidate the pathogenesis of ALV.