Solubilization of Membrane-Associated Cross-Protection Factor(s) of Pasteurella multocida

Abstract

Pasteurella multocida harvested from the blood of turkeys dying of experimental fowl cholera were purified by centrifugation and lysed. The soluble and membrane-associated components of the bacteria were separated by centrifugation. Nonionic (octylglucoside) and zwitterionic (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate; CHAPS) detergents were tested for their abilities to solubilize the cross-protection factor(s) (CPF) from the membrane-associated component. Protection studies in turkeys showed that optimum solubilization was by 1.0% octylglucoside and 0.5% CHAPS. Antibodies from turkeys made against solubilized membrane-associated CPF passively cross-protected poults against challenge. Ion exchange chromatography of detergent-solubilized CPF resulted in elution of two protein-containing peaks, each of which conferred active immune protection.