PREPARATION OF MONOCLONAL-ANTIBODIES FOR THE DIAGNOSIS OF INFLUENZA-AINFECTION USING DIFFERENT IMMUNIZATION PROTOCOLS

Four immunization protocols were used to obtain cross-reactive influen za type A-specific monoclonal antibodies: (1) repeated administration of purified influenza virus, (2) immunization with bromelain-treated v iral particles free of HA and NA, (3) sequential immunization with two strains of different subtypes, and (4) immunization with bromelain-tr eated particles following tolerization of mice to surface glycoprotein s by cyclophosphamide. The fourth approach was shown to be the most ef fective since a high proportion of hybridomas producing cross-reactive influenza virus type A-specific MAbs were obtained. MAbs of type A sp ecificity were immunochemically characterized and examined for their a bility to detect virus in clinical specimens. It was demonstrated that two pairs of the newly prepared MAbs provided excellent reagents for viral detection in clinical specimens using time-resolved fluoroimmuno assay.