Purification of Poliomyelitis Virus by Adsorption and Elution. ∗

Abstract

The exact nature of filterable viruses is considerably obscured by their close association with substances derived from the tissues in which the virus is found. The purpose of this communication is to describe a method whereby the virus of poliomyelitis may be partially purified, and ultimately perhaps be obtained in its pure state. The method follows closely the procedures which enabled Willstatter and his coworkers to isolate enzymes in their purest known form. Numerous investigators 1 have reported the capacity of various suspensions to adsorb filterable viruses. The adsorbed viruses are in most instances inactivated, and in the case of poliomyelitis, Amoss 2 states that “the presence of colloidal substances with adsorptive power destroys the virulence after 1 or 2 days.” Rhoads 3 described the adsorption and inactivation of poliomyelitis virus by aluminum hydroxide, type “C” of the Willstatter series, and showed that adsorption occurred at pH 5.5 and 7.0, but not at _p H 8.8. Gildemeister and Herzberg 4 succeeded in adsorbing bacteriophage on kieselguhr and subsequently eluting it with dilute ammonia. Kligler and Olitzki 5 confirmed these observations on bacteriophage, and were able to do the same with fowl-pox virus, using kaolin as the adsorbing agent.

If the inactivation of the poliomyelitis virus is not irreversible, it should be possible to adsorb it at an acid pH, and elute at an alkaline pH. For adsorption of the virus I used alumina gel “C”, prepared according to the method of Willstatter and Kraut 6 excepting that the centrifuge was used, instead of natural sedimentation. An effective gel was thus obtained, the process requiring only 2 days as compared with 2 weeks or more in the original procedure. The gel was standardized to contain 22-25 mg. Al₂ O₃ per cc.