Thiophilic adsorbents for RIA and ELISA procedures.
Three types of agarose derivatives have been prepared and investigated as adsorbents for radioimmunoassay and ELISA analysis. The analytical systems were evaluated using beta 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 x 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.
Biochemical Separation Centre, Uppsala University, Sweden.
S
S
Oscarsson
G
G
Chaga
J
J
Porath
D000818:Animals; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D008024:Ligands; D011725:Pyridines; D011863:Radioimmunoassay / Q000379:methods*; D012685:Sepharose / Q000031:analogs & derivatives*; D012756:Sheep; D001613:beta 2-Microglobulin / Q000032:analysis
D007074:Immunoglobulin G; D008024:Ligands; D011725:Pyridines; D001613:beta 2-Microglobulin; C071668:3-(N-pyridyl-2-one)-2-hydroxypropylagarose; C071667:3-(2-pyridylthio)-2-hydroxypropylagarose; D012685:Sepharose; C071669:3-(phenyloxy)-2-hydroxypropylagarose