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Thiophilic adsorbents for RIA and ELISA procedures

Authors
Journal
Journal of Immunological Methods
0022-1759
Publisher
Elsevier
Publication Date
Volume
143
Issue
2
Identifiers
DOI: 10.1016/0022-1759(91)90039-i
Keywords
  • Radioimmunoassay
  • Elisa
  • Thiophilic Adsorption
  • Immune Complex
Disciplines
  • Biology

Abstract

Abstract Three types of agarose derivatives have been prepared and investigated as adsorbents for radioimmunoassay and ELISA analysis. The analytical systems were evaluated using β 2 microglobulin as a model. After a competitive reaction between the immunocomponents in solution, the formed immune complexes were adsorbed onto the adsorbent in the presence of 0.5 M potassium sulfate in 0.1 M Tris, pH 7.5. The binding constant between the interaction site on human IgG and the adsorbent 3-(2-pyridylthio)-2-hydroxypropylagarose (Py-S-gel) was determined to be 1.5 × 10 7 M −1 and the binding capacity was 20 mg/ml gel. The immune complex was desorbed by deleting potassium sulfate from the buffer, and only 0.5% of the total applied protein remained after washing the adsorbent with 0.5 M NaOH. The same adsorbent can be used repetitively with different systems.

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