LARGE-SCALE PREPARATIONS OF VIRUSES BY STERIC CHROMATOGRAPHY ON COLUMNS OF CONTROLLED PORE GLASS - PHIX174, M13, M12, QBETA AND T4 BACTERIOPHAGES

Several bacteriophages of enterobacteria are quickly and effectively purified from highly concentrated crude extracts using columns of controlled pore glass without detectable loss of infectivity. The nucleic acids of the host cells are degraded by nucleases, and the bacterial debris is sedimented by low-speed centrifugation prior to column chromatography. Highly concentrated solutions of purified bacteriophages are eluted in the exclusion volume of the column. The preparation of the controlled pore glass columns and principles of operation are described. In the case of the Escherichia coli C bacteriophage ØX174, an exhaustive investigation of purity of glass-column chromatographed phage fractions has been made. For the other bacteriophages, sufficient evidence is presented to conclude that they will be as effectively purified as ØX174. The chromatographic step successfully removes both bacterial nucleases and the pancreatic nucleases used for degradation of the bacterial nucleic acids. © 1969.