ABSTRACT
This chapter explores the determination of the immunological activity of Sendai virus integral membrane proteins following size-exclusion high-performance liquid chromatography in different nonionic detergents using Mabs directed against the intact proteins. Polyclonal or monoclonal antibodies directed against linear epitopes may recognize the unfolded or denatured protein as well as the native protein. Immunological activity profiles were determined using Mabs directed exclusively against the native conformation of Sendai virus F and HN protein in an enzyme-linked immunosorbent assay. High-performance liquid chromatography (HPLC)-purified proteins may be used to immunize experimental animals to produce antibodies against the proteins. When the antiserum reacts with the intact protein or, in the case of viral proteins, is able to neutralize viral infectivity, this implies that a considerable part of intact structure must have been present, either consisting of linear or conformational epitopes. Depending on how the HPLC-purified protein will be used, a method should be selected to determine immunological activity.
