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

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

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Delphine H. ClarkeThe Yale Arbovirus Research Unit, Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, Connecticut

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Summary

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.

Author Notes

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.

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