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The Use of Sonication and of Calcium-Phosphate Chromatography for Preparation of Group C Arbovirus Hemagglutinins

The Yale Arbovirus Research Unit, Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, Connecticut

Various procedures were tried for improving the quality of group C arbovirus hemagglutinins derived from suckling-mouse liver. After separation of hemagglutinating (HA) and complement-fixing antigens by centrifugation, liver preparations were still too insensitive for use in hemagglutination-inhibition (HI) tests. Sonication produced marked improvement in HA titer, with no evidence that this was due to splitting of the virus particle; however, HI reactivity was still inadequate. Subsequent adsorption onto calcium phosphate (brushite form) followed by stepwise elution resulted in antigens of high quality.

Modified methods are described for preparation of sucrose-acetone antigens and for acetone extraction of immune mouse ascitic fluids.

^* Present address: Faculté de Médecine de Paris, Institut de Parasitologie, 15, rue de l'Écolede-Médecine, Paris, VI, France.

The Yale Arbovirus Research Unit is supported in part by The Rockefeller Foundation.