Capture and detection of carcinoembryonic antigen on antibody coated beads used in enzyme immunoassay.
We have evaluated antibody coated beads for capture and detection of carcinoembryonic antigen (CEA). Assay parameters of time, temperature, buffer molarity, specificity of antibody on the bead and reagent addition sequence have been studied. Optimal assay kinetics occurred at a temperature of 45 degrees C and a buffer molarity of 0.1M or above. The type and quantity of antibody on the bead surface were also critical to optimal CEA detection. Beads coated with baboon or goat anti-CEA antibody were able to capture a higher percentage of CEA than monoclonal mouse anti-CEA antibody or guinea pig anti-CEA antibody. The sequence of addition of CEA, anti-CEA antibody coated bead, and anti-CEA-horse radish peroxidase conjugate was important for optimal CEA detection. Formation of an immune complex of CEA with the anti-CEA horse radish peroxidase conjugate prior to capture of the CEA on an antibody bead resulted in the optimal detection of CEA.
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002272:Carcinoembryonic Antigen / Q000032:analysis* / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D007124:Immunoenzyme Techniques; D007700:Kinetics
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002272:Carcinoembryonic Antigen