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Purification of Concentrated Antitoxins

From the Biological Laboratories of E. R. Squibb and Sons, New Brunswick, New Jersey

Abstract

1. A study has been made of the application of the isoelectric method for further purification of concentrated antitoxins of varying age and previous treatment dialyzed to different specific conductances. The effectiveness of this method for removal of objectionable impurities was compared with that of fractional precipitation by salting out.

2. The potency and amount of protein found in the isoelectric precipitates varied not only with age and previous treatment of the antitoxin, but also with the conductivity of the dialyzed solution. The specific conductance of the diluted antitoxin solutions which afforded optimum practical separations was found to be between 125 and 85 x 10^-6.

3. In the concentration of antitoxins, heating of the plasma renders subsequent isoelectric purification of the concentrates more effective for the removal of phosphorus bearing materials.

4. After the removal of isoelectric protein from tetanus and diphtheria antitoxins and especially after electrodialysis the potencies of the fractions precipitated at lower concentrations of ammonium sulphate were increased. In the case of electrodialyzed tetanus the fractions precipitating between 34 and 43 per cent of saturation with ammonium sulphate, representing the bulk of the active protein, were of uniform potency.

5. Data are submitted which show quantitatively the superiority of the isoelectric method over salt fractionation for removing euglobulin, fat and phospholipins from concentrated antitoxin.