Quantification of Influenza Virus Structural Proteins using Rocket Immunoelectrophoresis

Abstract

The major influenza A virus structural antigens, matrix protein, nucleoprotein, hemagglutinin and neuraminidase were measured rapidly and accurately using a rocket immunoelectrophoresis technique. Virus was disrupted with 1% (wt/vol) sodium sarcosyl and electrophoresed into agarose containing specific antiserum to the individual virus structural proteins in 0.05 M-barbitone buffer, pH 8.6, for 1-4 h. For hemagglutinin antigen assays statistical analysis indicated that the coefficient of variation within an immunoelectrophoresis plate was 8.0% for antigen concentrations in the range 15-40 .mu.g/ml protein. For hemagglutinin and matrix protein the method was sufficiently sensitive to measure concentrations of antigens as low as 1.5 and 2.0 .mu.g/ml, respectively. By incorporation in the agarose of mixtures of antisera against specific antigens of the virus, hemagglutinin, matrix or nucleoprotein could be assayed simultaneously.