抗鸡IBDV Gt株单克隆抗体制备及抗原表位初步分析

Preparation and primary analysis of antigenic epitopes to monoclonal antibodies against Gt strain of chicken infectious bursal disease virus

采用不连续蔗糖密度梯度离心法纯化的鸡传染性法氏囊病病毒Gt株免疫BALB／C小鼠，利用淋巴细胞杂交瘤技术，取其脾细胞与SP2／0骨髓瘤细胞进行融合，经过三次筛选克隆，获得具有分泌抗IBDV抗体的7D4、5D2两株杂交瘤细胞系，并对7D4及5D2株进行了特异性、稳定性等方面的鉴定。试验表明，两株分泌的抗体能够特异地与IBDV VP2蛋白反应。7D4株及5D2株杂交瘤细胞上清ELISA效价分别为1：250、1：50，腹水ELISA效价为6．4×10^6、5×10^3；并运用7D4、5D2介导的相加ELISA进行抗原表位的初步定位，这两株单抗针对不同的VP2抗原表位。

The chicken infectious bursal disease virus of Gt strain was purified by discontinuous gradient ultracentrifugation. By using the hybridoma technique and three times cloning, 7D4 and 5D2 hybridoma cells secreting monoclonal antibody were established by the fusion of mouse myeloma cells SP2/0 and spleen cells from BALB/c mice immunized with purified IBDV. Then McAb was identified at isomerism and stabilitys, etc. Experimentation indicated that monoclonal antibodies have special reaction with the VP2 protein of IBDV. By indirect ELISA, the antibody titers of 5D2 and 7D4 were 2.5×10^2, 5×10^1 in culture supernatants and 6.4× 10^6, 5 ×10^3 in ascetic fluids. With sandwich ELISA the result of the antigen epitopes on different position of VP2 were basis located.