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Purification and Characterization of Coliphage MS2 Hapten Conjugates^1,2,

From the Department of Microbiology, University of Washington School of Medicine, Seattle, Washington and the Departments of Microbiology and Immunology, Mayo Graduate School of Medicine, Mayo Foundation, Rochester, Minnesota

Abstract

Conjugation of the RNA coliphage MS2 to either dinitrophenol (DNP) or picolinimidate (P), and separation from the reaction mixture on Sephadex G-25 results in the generation of at least two phage populations with differing sedimentation rates. These populations also can be separated on the basis of density. One phage population contains intact, infectious phage particles and conjugated hapten. The data presented shows that neither the density nor sedimentation rate change as a result of conjugation, nor is there any detectable alteration in the purified, conjugated particle that makes it any more sensitive to altered physical and chemical conditions. However, distinct differences between conjugated and unconjugated phage can be seen when they are neutralized with anti-MS2 sera.

Footnotes

¹ This investigation was supported by National Institutes of Health Fellowship Grant GM-49268 to Linda K. Curtiss from the National Institutes of General Medical Sciences and Public Health Research Grant AI-08044 from the National Institutes of Allergy and Infectious Diseases.

² Requests for reprints should be sent to: Department of Microbiology and Immunology, Mayo Graduate School of Medicine, Mayo Foundation, Rochester, Minnesota 55901.

³ Department of Microbiology, University of Washington School of Medicine, Seattle, Washington 98105.

⁴ Departments of Microbiology and Immunology, Mayo Graduate School of Medicine, Mayo Foundation, Rochester, Minnesota 55901.