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Published on: December 14, 2016
Preparation of monoclonal antibodies for the diagnosis of influenza A infection using different immunization
E Varecková1, T Betáková, V Mucha
1Institute of Virology, Slovak Academy of Sciences, Bratislava.
Four immunization protocols were used to obtain cross-reactive influenza type A-specific monoclonal antibodies: (1) repeated administration of purified influenza virus, (2) immunization with bromelain-treated viral particles free of HA and NA, (3) sequential immunization with two strains of different subtypes, and (4) immunization with bromelain-treated particles following tolerization of mice to surface glycoproteins by cyclophosphamide. The fourth approach was shown to be the most effective since a high proportion of hybridomas producing cross-reactive influenza virus type A-specific MAbs were obtained. MAbs of type A specificity were immunochemically characterized and examined for their ability 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 fluoroimmunoassay.
Four immunization protocols were used to obtain cross-reactive influenza type A-specific monoclonal antibodies: (1) repeated administration of purified influenza virus, (2) immunization with bromelain-treated viral particles free of HA and NA, (3) sequential immunization with two strains of different subtypes, and (4) immunization with bromelain-treated particles following tolerization of mice to surface glycoproteins by cyclophosphamide. The fourth approach was shown to be the most effective since a high proportion of hybridomas producing cross-reactive influenza virus type A-specific MAbs were obtained. MAbs of type A specificity were immunochemically characterized and examined for their ability 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 fluoroimmunoassay.
