Immunoelectrophoresis employing avian antisera for the detection and quantitation of Pasteurella multocida antigens.
Immunoelectrophoresis with various buffer systems at high and low pH was examined for suitability to detect and quantitate Pasteurella multocida antigens with turkey or chicken anti-P. multocida sera. Counterimmunoelectrophoresis was used to develop a buffer system for one-dimensional, two-dimensional, and rocket immunoelectrophoresis. The effects of pH, buffer, and molarity on resolution of immunoprecipitates were determined; 0.05 M sodium acetate-acetic acid buffer at pH 5.6 was the most suitable buffer. This buffer could be used in counterimmunoelectrophoresis with turkey or chicken sera to detect minute amounts of P. multocida protein antigens (4.3 ng/test) or lipopolysaccharide (3.12 micrograms/test). One-dimensional immunoelectrophoresis with the acetate buffer system required treatment of the gels with a 17% NaCl solution to induce immunoprecipitation of P. multocida lipopolysaccharide. Other techniques using the acetate buffer system did not require the high salt treatment. In two-dimensional immunoelectrophoresis, antisera migrated in the second dimension at pH 8.6, but did not migrate at pH 5.6. Rocket immunoelectrophoresis with the acetate buffer system was effective for quantitating P. multocida antigens.
D000818:Animals; D000906:Antibodies; D000942:Antigens, Bacterial / Q000032:analysis*; D002021:Buffers; D002645:Chickens / Q000276:immunology; D003378:Counterimmunoelectrophoresis / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007122:Immunoelectrophoresis / Q000379:methods*; D007123:Immunoelectrophoresis, Two-Dimensional / Q000379:methods; D008070:Lipopolysaccharides / Q000276:immunology; D009994:Osmolar Concentration; D010325:Pasteurella / Q000276:immunology*; D014422:Turkeys / Q000276:immunology
D000906:Antibodies; D000942:Antigens, Bacterial; D002021:Buffers; D008070:Lipopolysaccharides