抗牛免疫球蛋白G单克隆抗体的制备及鉴定

采用杂交瘤技术从90株融合株中筛选得到分泌抗牛免疫球蛋白G(immunoglobulin G,IgG)单克隆抗体的2F6、5E3、9C6、9H8四个细胞株。其抗体亚型均为IgG1型,腹水单克隆抗体的效价可达5.12×10~6;5E3、9H8识别牛IgG重链Fc片段,2F6识别牛IgG轻链Fab片段,9C6识别牛IgG重链Fab片段;9C6与牛IgG1和牛IgG2反应,与山羊IgG、绵羊IgG、兔IgG、牛IgM、牛乳酪蛋白、牛乳β-乳球蛋白、牛乳铁蛋白、牛血清白蛋白、鸡卵清白蛋白、鱼明胶的交叉反应率均小于1%;9C6亲合力常数达牛乳8.92×10~8 L/mol。

Preparation and Identification of Monoclonal Antibody against Bovine Immonoglobulin G

Four hybridomas secreting monoclonal antibody (McAb) against bovine immunoglobulin G (IgG) were developed by fusion of spleen cells from immunized BALB/C mice with SP2/0 myeloma cells, and they were named as 2F6, 5E3, 9C6 and 9H8. The McAbs secreted by all four hybridomas were identified as IgG1 subtype with a titer of 5.12 × 106. Both 5E3 and 9H8 recognized the heavy chain Fc fragment of bovine IgG. 2F6 recognized the light chain Fab fragment of bovine IgG, and 9C6 recognized the heavy chain Fab fragment of bovine IgG. The McAb from 9C6 could react with both IgG1 and IgG2. The cross-reactivity between 9C6 and goat IgG, sheep IgG, rabbit IgG, bovine IgM, casein, β-lacto globulin, lactoferrin, bovine serum albumin, ovalbumin or fish gel was less than 1%. The affinity constant of 9C6 was 8.92 × 108 L/mol.