A comparison of fluorescein-labeled gamma-globulins purified by Rivanol and DEAE chromatography

Preparations of fluorescein-labeled γ-globulins obtained by the Rivanol procedure were found to be comparable to DEAE γ-globulins in recovery of antibody and specificity of fluorescent staining. An application of these preparations was made to the fluorescent antibody analysis of the V antigens of influenza virus where it was necessary to use purified, labeled γ-globulins to achieve full specificity. However, in other virus-cell systems immune globulins prepared by ammonium sulfate precipitation, conjugated at low F:P ratios, and passed through a sephadex column were found to be comparable to the Rivanol and DEAE globulins in specificity and, at times, superior in stainability. In the reverse situation, where essentially no contaminating host materials are present, as with preparations of bacteria, the use of ammonium sulfate globulins conjugated at high F:P ratios seems to afford the greatest intensity of staining.