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Rubella Complement-Fixing Antigens Derived from the Fluid and Cellular Phases of Infected BHK-21 Cells: Extraction of Cell-Associated Antigen with Alkaline Buffers¹

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

Abstract

Rubella complement-fixing (CF) antigens with titers considerably higher than those previously prepared from infected RK-13 culture fluids were obtained by concentration of the fluid phase of infected BHK-21 cultures. Greater amounts of CF antigen could be obtained from the cellular phase of infected BHK-21 cultures by sonic oscillation than by freezing and thawing, and extraction of infected cells with glycine or borate buffers at pH 9 or 10 released even more antigen. In addition to being more potent antigens, alkaline buffer extracts were less anticomplementary than antigens produced by physical disruption of the cells. Factors influencing the effectiveness of extraction of CF antigen with alkaline buffers were studied. Antigens produced from both the fluid and cellular phases of infected BHK-21 cultures could be treated with ether to render them noninfectious with only a slight loss in titer; ether-treated antigens retained their sensitivity and specificity in tests with human sera. The preparation of both fluid concentrates and alkaline buffer extracts of infected cells greatly increased the amount of rubella CF antigen obtainable from infected BHK-21 cultures.

Footnotes

¹ The work on which this paper is based was supported by Grant AI-01475-06 from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Public Health Service, Department of Health, Education and Welfare.