Studies on Tetanus Toxin and Toxoid I. Isolation of Tetanus Toxin using Deae-Cellulose

Abstract

Tetanus toxin has been extracted from culture filtrates by a single, cold methanol precipitation, followed by gradient elution on DEAE-cellulose. Immunoelectrophoretic evidence indicated that most of the bacterial protein was precipitated by the methanol. Selection of the toxin elution peak gave a product having only one immunoelectrophoresis precipitin line yet containing 50--60% of the Lf units applied to the column. The toxicity of this product was comparable to that found by other workers, using other methods of preparation. Sedimentation values of the concentrated pure toxin indicated heterogeneity in molecular aggregation and under conditions used no 4 S material could be found. Amino acid analysis of the tetanus protein is also reported.

Tetanus Toxin and Antigenic Derivatives I. Purification of the Biologically Active Monomer

“…The properties of tetanus toxin isolated by our procedure are similar to those reported by other investigators (1,18). It is difficult to compare Lf values due to the different reference standards employed by other investigators.…”

“…8. Agar double-diffusioni pattern of CEX (1) and pure toxin (2), 500 Lf/ml. Horse anticrude tetanus antibody was employed at a concentration of 500 Lf/ml.…”

“…Recently, several chromatographic purifications have been reported. For these procedures, either ion-exchange chromatography (1,8,15,17) or gel filtration (7, 15) has been used. None of the preparations obtained by these methods has been shown to be a single component with respect to both immunological and chemical criteria of purity.…”

Tables of Toxicity of Botulinum and Tetanus Neurotoxins

“…Tetanus toxin is most potent when introduced into the central nervous system, and least potent by mouth; the toxicity by oral route being 1/200,000-1/1,200,000 that of the parenteral route [83]. [17,47,81,[84][85][86]; data have been converted into nanograms/kilograms for a better comparison. b TeNT toxicity value for humans is the results of extrapolation from monkey data [6] and from records of accidental inoculations quoted by [47].…”

Tetanus toxin.

“…In the last two decades, the toxin has been isolated in a homogeneous state (12,21,22,52,65,73), and its physical and chemical characteristics have been defined in various laboratories in different parts of the world (5,12,14,20,22,50,52,63,64,73,83,85; S. G. Murphy, T. H. Plummer, and K. D. Miller, Fed. Proc., p. 268, 1968).…”