Amino Acid analysis and molecular weight determination of tetanus toxin
1Departments of Biochemistry and Obstetrics and Gynecology, University of Nebraska College of Medicine, Omaha, Nebraska 68105.
Abstract:
A procedure for the isolation of tetanus toxin was developed. The toxin was precipitated with ammonium sulfate and further purified by use of molecular sieve chromatography and preparative disc gel electrophoresis. The degree of purification resulting from these fractionation procedures was monitored by analytical poly-acrylamide gel electrophoresis, immunodiffusion, and immunoelectrophoresis, all three of which demonstrated a single component. There was a 50-fold increase in specific activity between the crude culture filtrate and the final purified product, which had an average specific activity of 300 x 10(6) mouse minimal lethal doses per mg of N. A molecular weight of 140,000 was estimated by gel filtration on Bio-Gel P-200. A complete amino acid analysis of tetanus toxin is presented.
Related Concept Videos
Tetanus
Mass Spectrometry of Amines
SDS-PAGE
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
Mass Spectrometry: Amine Fragmentation
In amines, the number of nitrogen atoms affects the mass of the molecular ion, which is described by the nitrogen rule of mass spectrometry. This rule states that a compound containing a single or...


