Studies on the Purification of Poliomyelitis Virus

Abstract

The application of high speed differential centrifugation to the purification of poliomyelitis virus has been a logical step since Stanley and Wyckoff (1) showed that relatively small and unstable plant viruses could be isolated as homogeneous materials by this method. The sedimentation of poliomyelitis virus in an ultracentrifuge was first demonstrated by Schultz and Raffel (2), and more recently Clark, Rasmussen, and White (3) obtained sediments which showed an irregular but in some instances high virus activity. While it has been shown that high speed centrifugation resulted in a sedimentation of the virus, evidence has not been provided that other macromolecular materials associated with normal tissue were not also sedimented along with the virus under these conditions. In view of the work of several investigators (4--6) on the concentration of macromolecular materials from various normal animal tissues, such constituents might very well be present together with virus in the high molecular weight fractions obtained from poliomyelitis-infected tissues.This paper presents the results of experiments on the application of differential centrifugation to the extracts of glycerolated or frozen, infected medullaecords from rhesus monkeys and to similar extracts of glycerolated or frozen, normal medullae-cords. The results show that when extracts of infected as well as normal cords, that have been stored in glycerol, are subjected to high speed centrifugation, appreciable yields of high molecular weight substances are obtained. However, when normal tissue that had been frozen and stored for appreciable lengths of time was used, no significant amount of high molecular weight, nitrogen-containing compounds was recovered. Frozen infected tissue, on the contrary, yielded small but significant amounts of high molecular weight material possessing both a high and uniform virus activity. EXPEB IM~.NTAL

Columbia SK Group of Viruses

“…Electron micrographic studies of this purified preparation by GoLLAN and MARVIN (1948) showed uniform particles in the range from 10-12 mf1, with an average diameter of about 12 IDf1. These findings are in agreement "\\<ith the early measurements of purified LANSING poliomyelitis virus by LORING et al (1942;. However, the question arises whether the observed particles were really the infectious unit or the smaller component which is currently thought to be associated with the complement-fixing property of polioinyelitis virus.…”