为制备抗鸡岱:微球蛋白(βz-M)单克隆抗体(MAb),本研究以合成的多肽TF-29作为免疫原免疫BALB/c小鼠,利用杂交瘤技术筛选出特异性的抗鸡p2-M杂交瘤细胞株(3D1和6E7),亚类鉴定均为IgG1/K链;间接EUsA测定3D1和6E7腹水效价分别为1:25600和1:204800。应用westernblot和间接免疫荧光鉴定MAb的特异性显示,其不仅能够与原核表达产物也能够与真核细胞表达的盼M特异性结合,还能够与禽类巨噬细胞系HD11中天然的β2-M特异性反应。3D1和6E7也能够特异性识别正常鸡血清与血浆中游离的β2-M。以上结果表明,本研究应用多肽筛选出抗鸡盼MMAb,其效价稳定并具有良好的特异性,为深入探索鸡BrM在免疫学中的功能提供实验依据。
Two monoclonal antibodies (MAbs) against chicken [32-microglobulin ([32-M) were prepared by fusions the SP2/0 cells with spleen cells from BALB/c mice immunized with a synthesized peptide (TF-29). The ELISA titers of two antibodies 3D1 and 6E7, identified as IgG1/K, were 1:25 600 and 1:204 800 respectively. Both MAbs specifically recognized not only the recombinant β2-M expressed in prokaryotic cells, but also the native chicken β2-M in avian macrophage HD11 cells by western blot and immunofluorescence assays. Moreover, both MAbs were able to react with the β2-M from chicken serum and plasma in western blot assay. These MAbs would facilitate for further study of chicken β2-M in the regulation of the immune system in chicken.