Skip to main content
Applied Microbiology logoLink to Applied Microbiology
. 1967 May;15(3):616–621. doi: 10.1128/am.15.3.616-621.1967

Preparative Procedure for the Purification of Toxoids by Gel Filtration

William C Latham 1,2, Christopher B Michelsen 1,2, Geoffrey Edsall 1,2
PMCID: PMC546987  PMID: 4962287

Abstract

Sephadex gel filtration can be employed as a preparative procedure for the purification of both tetanus and diphtheria toxoids. A toxoid purification sequence is described in the text. By utilizing the described methods and columns, up to 100,000 human doses of diphtheria toxoid could be processed in a single operation. The method has given an 80% yield of diphtheria toxoid with a purity of 1,900 Lf per mg of N. The analysis of the material by immunodiffusion tests showed that a marked increase in purity was achieved. Antigenicity tests demonstrated that there was no significant difference in antigenic potency between the parent toxoid and its purified fraction. Factors limiting the effective separation of tetanus toxoid by gel filtration are discussed. The construction of the columns used is described in detail, as well as packing procedures and column characteristics such as bed volume, void volume, sample size, and flow rate.

Full text

PDF
617

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. LEVINE L., STONE J. L. The purification of tetanus toxoid by ammonium sulfate fractionation. J Immunol. 1951 Sep;67(3):235–242. [PubMed] [Google Scholar]
  2. LEVINE L., WYMAN L., EDSALL G. Chemical and immunological properties of diphtheria toxoid purified by ammonium sulfate fractionation. J Immunol. 1949 Nov;63(3):219–232. [PubMed] [Google Scholar]
  3. Latham W. C., Jenness C. P., Timperi R. J., Michelsen C. B., Zipilivan E. M., Edsall G., Ley H. L., Jr Purification and characterization of tetanus toxoid and toxin. I. Fractionation of tetanus toxoid by gel filtration. J Immunol. 1965 Sep;95(3):487–493. [PubMed] [Google Scholar]
  4. PORATH J., BENNICH H. Recycling chromatography. Arch Biochem Biophys. 1962 Sep;Suppl 1:152–156. [PubMed] [Google Scholar]
  5. Robb L. A. Separation of diphtheria toxin proteins by Sephadex gel. Can J Microbiol. 1966 Jun;12(3):573–579. doi: 10.1139/m66-080. [DOI] [PubMed] [Google Scholar]

Articles from Applied Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES