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A Comparison of Fluorescein-Labeled -Globulins Purified by Rivanol and Deae Chromatography¹

From the Viral and Rickettsial Disease Laboratory, California State Department of Public Health, Berkeley, California

Abstract

Preparations of fluorescein-labeled -globulins obtained by the Rivanol procedure were found to be comparable to DEAE -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 -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.

Footnotes

¹ The work on which these observations are based was supported by Grant E-1475 from the National Institute of Allergy and Infectious Diseases, United States Public Health Service, Department of Health, Education and Welfare.