A Chromatographic Procedure for the Purification of Influenza Virus

Abstract

SUMMARYOwing to the difficulties of achieving consistently good recoveries of virus from allantoic fluid containing influenza virus by chromatography on calcium phosphate columns the factors determining optimum yield were systematically investigated. It was found that calcium phosphate prepared by precipitation at a controlled pH of 7.5 gave best results provided that eluting buffers were adjusted to the same pH. Although calcium phosphate fractions were still evidently contaminated with non-viral material, further purification could be obtained by chromatography in agarose-gel columns. It appeared that separation of the virus on these columns was probably achieved as a result of the binding of virus surface groups on to the gel. A minimum overall purification factor of about 800-fold was achieved.

Antigenic properties of the envelope of influenza virus rendered soluble by surfactant-solvent systems

“…The virus pellet was taken up in a volume of BS or de-ionized water equivalent to the original volume of the eluent (10 % Na2HPO4). Since the determination of an overall purification factor can be hampered by the extreme lability of highly concentrated virus preparations (Pepper, 1967), the calculations to determine purification factors were made on HA titres immediately before and after purification.…”

Substituted Sialic Acid Prosthetic Groups as Determinants of Viral Hemagglutination

“…The two strains of influenza virus used in this study were the inhibitor-sensitive variant of A2/Singapore/57 (5) previously used in this department and B/England/13/65, originally received from A. S. Beare. The viruses were grown and purified as already described (14).…”

“…Molecular exclusion chromatography. A column of agarose beads (4% gel, 120 mesh, 1.16 by 123 cm) was prepared and calibrated (14). The sample was layered on the column and eluted at 3.0 ml/hr with 0.15 M NaCl containing 0.02% NaN3.…”