Purification and characterization of IgG1 and IgG2 from buffalo (Bubalus bubalis) serum and colostrum.
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.
Kakker
N K
NK
Department of Veterinary Microbiology, Haryana Agricultural University, Hisar, India.
Goel
M C
MC
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
D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains