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Factors Influencing the Rate of Neutralization of Bacteriophage by the Antibody

From the Department of Bacteriology and Immunology, Washington University School of Medicine, St. Louis, Missouri

Abstract

1. Employing an antiserum prepared against a highly purified preparation of coliphage, we have been able to demonstrate that the specific rate of neutralization of phage by antibody is independent of the absolute phage concentration, that is, a constant proportion of phage is neutralized in a given time regardless of what concentration of phage is actually present.

2. The course of neutralization progresses logarithmically with time, that is, a constant fraction of phage is neutralized in each successive time interval. Deviations are observed only when the reaction is allowed to go to better than 99 per cent of completion.

3. The neutralization of phage by antiphage resembles a (bimolecular) reaction in which two molecules of antibody are needed to neutralize one particle of phage.

4. The logarithm of the proportion of phage remaining active after a given time-interval is proportional to the serum-concentration except in the case of very concentrated serum where more phage remains unneutralized than would be expected.

5. These results may be explained by assuming either that phage suspension consists of particles heterogeneous in their resistance to inactivation by serum, as suggested by Andrewes and Elford, or else by assuming that the bulk of the phage is relatively homogenous, but contains a small fraction (less than 1%) that is arkedly more resistant to neutralization.

6. The reaction of neutralization of phage at 37 C. proceeds at a rate 12 to 14 times faster than at 0 C. (has a Q₁₀ of 2 from 0 to 37 C.)

7. The reaction is relatively indifferent to pH except that below pH 4.5 the rate of enutralization is markedly slowed.

8. Identity of rates of neutralization of small and large particles indicate the probable correctness of the theory that phage of different sizes consists of small active molecules adsorbed to inert carriers of different sizes.