Affinity purification and partial characterisation of systemic immunoglobulin of the snapper (Pagrus auratus)

摘要: Immunoglobulins (Ig) from tank housed wild caught snapper (Pagrus auratus, Bloch and Schneider) were single step purified serum using staphylococcal protein A (SpA) affinity chromatography. Purified proteins analysed SDS-PAGE under reducing, non-reducing denaturing PAGE native conditions. Under conditions, a population of Ig was identified gel filtration chromatography found to have an approximate molecular weight 766 kDa. Further, non-denaturing reducing conditions the be heterogeneous in subunit linkages. subjected fully dissociated into heavy (H) light (L) chain polypeptides. Two H two L variants based differences electrophoretic mobility detected by SDS-PAGE, however evidence isotypic disparity not proven. The chains shown approximately 30.2 29.0 kDa while 71.8 67.7 kDa, suggesting that molecule likely tetrameric structure. Polyclonal antisera against produced screened indirect ELISA, Western blot flow cytometry. Specificity demonstrated probing Ig, whole heterologous blots. Antisera reacted predominantly with both reduced serum. lack cross-reactivity five six sera tested, high degree specificity antiserum. In cytometry, antiserum bound putative B cell population, reacting 40.8% (rabbit 1) 29.5% 2) gated lymphocytes peripheral blood.