抗鸡免疫球蛋白(IgM、IgG)单克隆抗体的制备与鉴定

Preparation and Identification of Monoclonal Antibodies Specific for Chicken IgM and IgG

本文报道将SRBC免疫鸡的血清经33％硫酸铵沉淀,再经Sephadex G-200凝胶柱两次层析,收集第Ⅰ,Ⅱ蛋白峰作为免疫原免疫BALB/c小鼠。建立了2株(3A7、1C4)分泌抗鸡IgG和4株(6AS、5E2、6C7、6B1)抗鸡IgM特异性单克隆抗体的杂交瘤细胞系;琼脂扩散试验表明6株细胞系分泌的单克隆抗体均为IgG1亚类。所制腹水的ELISA滴度达10~5～10~6。用免疫印迹技术(IB,Immunobllotting)对其中2株(3A7和6A5)的特异性作了进一步鉴定。

Sera from chickens immunized with SRBC were precipitated with 33% (HN4)2SO4, followed by filtration through sephadex G-200 gel column chromatogra-phy. The first and second protein peaks were collected as immunogens of chicken IgM and IgG, respectively, and they were used to immunize BALB/c mice. Two .hybridoma. cell lines ( 3A7, lC4 ) secreting McAbs against chicken IgG and four hybridoma cell lines ( 6A5, 6Cr, 6Bl, and 5E2 ) secreting McAbs specific for chicken IgM were obtained. All of the McAbs fell into the subclass of IgG1 as determined by double gel diffusion test. The tit'ers of ascitic fluid containing these McAbs were 10-4-10-4 by ELISA. The specificity of the McAbs ( 3A7 and 6A5 ) was further identified by imm-unoblotting.