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J Virol. 1968 September; 2(9): 937-943
Copyright © 1968 American Society for Microbiology. All Rights Reserved.

Use of Zonal Ultracentrifuge Systems for Biophysical Studies of Rhinoviruses

Department of Infectious Disease and Pathology, Scientific Division, Abbott Laboratories, North Chicago, Illinois 60064

ABSTRACT

This paper reports the use of zonal ultracentrifuge techniques to conduct biophysical studies of rhinoviruses grown with WI-38 cells. Good clean-out of infectivity from rhinovirus harvests was obtained with the continuous-flow B-V and B-IX rotors. Use of the B-V rotor resulted in the successful concentration of rhinovirus infectivity and antigenicity. Additional purification was achieved by the combined use of continuous-flow centrifugation and isopycnic banding procedures. Two particle sizes were found to be associated with the virus-infected cell harvests. The infectious 22-nm particle banded in density ranges of 1.38 to 1.40 g/cm³ in CsCl and 1.26 to 1.27 g/cm³ in potassium citrate. The 8.0 nm capsomere was composed of 2.0 nm subunits and banded with a density of protein at 1.28 g/cm³ in CsCl. Equivalent sedimentation coefficients of 155 or 185, depending on particle density in sucrose, were calculated from rate zonal experiments by use of the B-IV zonal rotor.

¹ Present address: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Building 7, Room 304, National Institutes of Health, Bethesda, Md. 20014.

² Present address: University of Chicago, Department of Pathology, Chicago, Ill. 60637.

³ Present address: Illinois Institute of Technology, Research Institute, Life Sciences, Chemistry Building, 10 West 35th Street, Chicago, Ill. 60616.