A sensitive enzyme immunoassay for the detection of a synthetic affinity ligand, the reactive yellow 13 dye.
Dye-affinity chromatography is widely and increasingly used for the isolation of various proteins. In particular, the purification of transthyretin can be efficiently achieved by affinity chromatography on immobilized Reactive Yellow 13. Measurement of trace-amounts of dye leaching from affinity columns is important because of possible toxicity or side effects linked with the presence of dye in therapeutic transthyretin preparations. A competitive enzyme immunoassay was developed to monitor yellow-dye column leaching. Biotinylated rabbit anti-Reactive Yellow 13 antibodies (immunoglobulin G fraction) were used as principal reagent. The assay is specific sensitive to 1 ng/ml of Reactive Yellow 13, has a good reproducibility and allows the accurate detection of the dye in the presence of transthyretin.
Regnault
V
V
INSERM U 284, Vandoeuvre-lès-Nancy, France.
Vallar
L
L
Rivat
C
C
Stoltz
J F
JF
Boschetti
E
E
D000345:Affinity Labels / Q000032:analysis; D000918:Antibody Specificity; D001391:Azo Compounds / Q000032:analysis* / Q000276:immunology; D002846:Chromatography, Affinity; D004396:Coloring Agents / Q000032:analysis*; D004340:Drug Contamination; D005069:Evaluation Studies as Topic; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D011228:Prealbumin / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D000345:Affinity Labels; D001391:Azo Compounds; D004396:Coloring Agents; D011228:Prealbumin; C035187:reactive yellow 13