Isolation and molecular weight determination of two immunoglobulin heavy chains in the channel catfish, Ictalurus punctatus.
Channel catfish (Ictalurus punctatus), a teleost fish, were immunized over a 4 month period with 4 intraperitoneal injections of bovine serum albumin (BSA) in Freund's adjuvant. The catfish anti-BSA antibody was purified by affinity chromatography and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). By elution of catfish anti-BSA antibody from BSA-affinity columns with 3.0 M KSCN and subsequent SDS-PAGE, two immunoglobulin heavy chains were demonstrated in the channel catfish. The molecular weights and the relative percentages found of the two immunoglobulin heavy chains were 72,000 (94%) and 56,000 (6%). The molecular weight of the single light chain found was 23,000. Using the 72,000 mol. wt heavy chain and 23,000 mol. wt light chain and including a molecular weight of 15,000 for the J-chain, the molecular weight of the predominant channel catfish tetrameric IgM immunoglobulin molecule was calculated to be 775,000. Using the 56,000 low mol. wt heavy chain, the molecular weight of a second subclass of the channel catfish tetrameric IgM molecule was calculated to be 647,000. After Sephadex G-200 gel filtration, anti-BSA antibody activity was found only in the 14S globulin fraction by indirect hemagglutination testing.
D000818:Animals; D002397:Catfishes / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D006386:Hemagglutination Tests; D007059:Ictaluridae / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000302:isolation & purification*; D008970:Molecular Weight
D007143:Immunoglobulin Heavy Chains