Original scientific paper

Fractionation of Tetanus Toxoid by Sephadex Gel Filtration and Immunochemical Characterization of the Fractions

M. Fleš ; Institute of Immunology, Zagreb, Croatia, Yugoslavia
B. Pende ; Institute of Immunology, Zagreb, Croatia, Yugoslavia

Abstract

Three different ty;pes of tetanus toxoid were fractionated on
Sephadex G - 200 gel. The distribution of the fractions was determined
spectrophotometrically at 280 mfL . Specific activity of the fractions
in vitro was determined by flocculation with standard tetanus antitoxin
and by means of immunodiffusion and immunoelectrophoresis.
In all three types of the toxoid the active component appeared in
the first three fractions ; about 750/o was found in the third fraction.
The purity of this fraction, in terms of Lf/mg. of protein nitrogen
(PN), was increased about twic e in comparison with the starting
material. The highest purity, 2400 Lf/ mg. PN, was achieved in the
third fraction of the toxoid type B-1, which w as p repurified, first
by ultrafiltration of the toxin, then, after detoxication, by fractional
precipitation with ammonium sulfate. This fraction was homogeneous
according to immunodiffusion and immunoelectrophoresis te sts.
It has been concluded that fractional purification of t etanus
toxoid can be effectively achieved by gel filtration on Sephadex
G-200, but the effect of purification highly depends on the purity
of the starting material.

Keywords

Hrčak ID:

208093

URI

https://hrcak.srce.hr/208093

Publication date:

10.8.1967.

Visits: 725 *