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Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
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Procedures for In Vitro Cultivation of Treponema pallidum, the Syphilis Spirochete
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BIOCHEMICAL STUDIES ON SO-CALLED SYPHILIS ANTIGEN.

H Noguchi1, J Bronfenbrenner

  • 1Laboratories of the Rockefeller Institute for Medical Research, New York.

The Journal of Experimental Medicine
|October 30, 2009
PubMed
Summary

The study identified that liver extracts contain various substances, with aceton-insoluble fractions being the most potent for antigen preparation in syphilis diagnosis. These fractions, rich in phosphatids, offer superior antigenic properties compared to other extract components.

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Area of Science:

  • Biochemistry
  • Immunology
  • Analytical Chemistry

Background:

  • Liver tissues contain diverse substances extractable by alcohol.
  • Antigenic properties are concentrated in specific fractions of these extracts.
  • Fractionation is crucial for isolating active components for diagnostic reagents.

Purpose of the Study:

  • To fractionate alcoholic liver extracts and identify components with antigenic properties.
  • To evaluate the biological activity and suitability of different fractions as antigens.
  • To recommend the optimal fraction for antigen preparation, particularly for the Wassermann reaction.

Main Methods:

  • Alcoholic extraction of liver tissues from humans and animals.
  • Fractionation of extracts based on solubility in ether, alcohol, and acetone.
  • Biological evaluation including hemolytic, anticomplementary, and antigenic property assessment.

Main Results:

  • Four distinct fractions were isolated based on solubility.
  • Fractions insoluble in ether and hot alcohol (proteins, salts) were unsuitable due to complement binding.
  • Fractions soluble in ether and alcohol (soaps, bile salts) were hemolytic and had weak antigenicity.
  • Fractions soluble in ether, alcohol, and acetone (lipoids) showed variable hemolytic and antigenic properties.
  • Fractions insoluble in acetone (phosphatids) demonstrated the most consistent and potent antigenic properties, despite occasional anticomplementary activity.

Conclusions:

  • The aceton-insoluble fraction, primarily containing phosphatids, is the most suitable for antigen preparation.
  • This fraction exhibits strong antigenic properties and is recommended for diagnostic use, such as in the Noguchi method for syphilis.
  • Further analysis of phosphatids within this fraction is warranted.