一种针对鹅免疫球蛋白轻链恒定区的新型单克隆抗体。
A novel monoclonal antibody against the constant region of goose immunoglobulin light chain.
作者信息
Guo Yongli, Gao Mingchun, Ma Bo, Sheng Qiaoling, Wang Qian, Liu Dandan, Wang Junwei
机构信息
Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University , Harbin, China .
出版信息
Monoclon Antib Immunodiagn Immunother. 2014 Apr;33(2):121-5. doi: 10.1089/mab.2013.0071.
A monoclonal antibody (MAb) against the antigenic determinant of the constant region of goose immunoglobulin light chain (GoIgCL) was produced and characterized for the first time here. Goose immunoglobulin (Ig) in serum was purified by immunoaffinity chromatography and the resulting protein was used as immunogen to immunize BALB/c mice. At the same time, the GoIgCL gene was expressed and purified as the screening antigen for selecting MAb against GoIgCL. One hybridoma that produces antibodies against GoIgCL was selected by indirect ELISA. Then the characterization of the MAb was analyzed by ELISA, Western blot, and flow cytometry. It was found to be IgG1 with κ light chain; the MAB has high specificity to Ig in goose serum, bile, and B lymphocytes from peripheral blood, reacts only with the light chain of goose Ig, and can distinguish Ig from other birds. Therefore, the MAb generated in this study can be used as a specific reagent for detection of goose disease-specific antibodies and as a powerful tool for basic immunology research on geese.
本文首次制备并鉴定了一种针对鹅免疫球蛋白轻链（GoIgCL）恒定区抗原决定簇的单克隆抗体（MAb）。血清中的鹅免疫球蛋白（Ig）通过免疫亲和层析法进行纯化，所得蛋白质用作免疫原免疫BALB/c小鼠。同时，表达并纯化GoIgCL基因作为筛选抗GoIgCL单克隆抗体的抗原。通过间接ELISA筛选出一株产生抗GoIgCL抗体的杂交瘤。然后通过ELISA、Western印迹和流式细胞术分析该单克隆抗体的特性。结果发现其为具有κ轻链的IgG1；该单克隆抗体对鹅血清、胆汁和外周血B淋巴细胞中的Ig具有高度特异性，仅与鹅Ig的轻链反应，且能区分鹅Ig与其他鸟类的Ig。因此，本研究中产生的单克隆抗体可作为检测鹅疾病特异性抗体的特异性试剂，以及鹅基础免疫学研究的有力工具。
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