The Getting, the Purification and the Characterization of Species’Immunoglobulin G (IgG) and Anti-immunoglobulin G Serum for Dog,Goat and Rabbit
Abstract
The research aimed at preparing, purifying and characterizing the immunoglobulin G, for dog, goat and rabbit, and the anti-IgG serum for the same species. The IgG species purification was done through precipitation, using ammonium sulfate solution and ion exchange chromatography (DEAE-cellulose). Checking the IgG purity was done by immunoelectrophoresis (IEF), for the corresponding anti-species serum, and by polyacrylamide gel electrophoresis in denaturant system (SDS-PAGE). Using IEF, we obtained a precipitation arch with cathodic migration, whereas using SDS-PAGE, we obtained a fraction with a molecular mass of 150 KDa. Dog and goat anti-IgG serum was prepared on rabbit, and the rabbit anti-IgG serum on goat, by IgG hyperimmunisation concealed in Freund adjuvant at a 1 to 1 ratio. Sensitivity and specificity testing of antisera was performed by immunoenzymatic test (ELISA). There were highlighted the high titers of antibodies to the dilution of 1:500 for anti-IgG rabbit and goat sera, and 1:400 for anti-IgG dog serum. Specificity testing of dog and rabbit anti-IgG in the heterologous system showed results which are in the negative range. The DO values were high for goat anti-IgG serum, sheep IgG serum and sheep anti-IgG serum and goat anti-IgG serum, due to antigenic kinship between these species. The reagents prepared will be used in great specificity and sensitivity immunodiagnostic tests.
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