RESUMO
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.
Assuntos
Anticorpos Monoclonais Murinos/química , Gansos/imunologia , Regiões Constantes de Imunoglobulina/imunologia , Cadeias Leves de Imunoglobulina/imunologia , Animais , Anticorpos Monoclonais Murinos/biossíntese , Especificidade de Anticorpos , Proteínas Aviárias/imunologia , Proteínas Aviárias/isolamento & purificação , Western Blotting , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Hibridomas , Regiões Constantes de Imunoglobulina/isolamento & purificação , Cadeias Leves de Imunoglobulina/isolamento & purificação , Linfócitos/imunologia , Camundongos Endogâmicos BALB CRESUMO
A novel gene encoding the α-chain of goose immunoglobulin heavy-chain (Igα) was cloned by reverse transcription-PCR. The cDNA had 1,760 bp and encompassed a partial V-D-J region of the heavy chain, a constant region (Cα) and 3'-untranslated region of α-chain. The Cα gene contains four constant region domains (CH1-CH4). Phylogenetic analysis indicated that goose IgCα has a close genetic relationship with duck, ostrich and chicken IgCα. Three-dimensional modeling and glycosylation analysis revealed the goose Igα is consistent with the characterization of immunoglobulin. Western blotting suggested the goose IgCα has the same antigenicity to natural IgA. In general, the identification of goose immunoglobulin not only provides insights into the evolution of the Ig heavy-chain gene family, but may also benefit future studies of the avian immune system.
