title: Purification and characterization of IgG1 and IgG2 from buffalo (Bubalus bubalis) serum and colostrum.
abstract: Buffalo IgG1 and IgG2 were purified from serum and colostrum using salt precipitation, dialysis, gel filtration and ion-exchange chromatography. Their purity was monitored by immunodiffusion and immunoelectrophoresis using anti-heavy chain specific sera and SDS-PAGE. Selective binding of IgG2 to protein-A was used to remove IgG2 from IgG1 preparations. The IgG1 and IgG2 had a molecular mass (Mr) of 162.0 and 161.5 kD, respectively and were found to consist of heavy (H) and light (L) chains. The H and L chains had Mr of 58 and 24 kD, respectively. Reduction-alkylation followed by gel filtration was used for the isolation of H and L chains. While intact H chains were obtained, the L chains appeared to be cleaved into 14 kD molecules and smaller fragments. The mean hexoses content of the serum IgG1 and IgG2 was 1.81 +/- 0.02% and 0.70 +/- 0.02%, respectively. The corresponding values for colostral IgG1 and IgG2 were 1.76 +/- 0.01% and 0.78 +/- 0.08%. Both the IgG subclasses activated homologous complement. These results suggest that buffalo and cattle IgG subclasses have many common characteristics and minor differences.
authors.affiliation: Department of Veterinary Microbiology, Haryana Agricultural University, Hisar, India.
authors.forename: N K
authors.initials: NK
authors.lastname: Kakker
authors.forename: M C
authors.initials: MC
authors.lastname: Goel
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000276:immunology*; D011232:Chemical Precipitation; D002850:Chromatography, Gel / Q000662:veterinary; D002852:Chromatography, Ion Exchange / Q000662:veterinary; D003126:Colostrum / Q000276:immunology*; D003168:Complement Fixation Tests; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification*; D007143:Immunoglobulin Heavy Chains / Q000276:immunology / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D011247:Pregnancy
chemical_list: D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains