Effects of supermolecule additions to the enzyme-linked immunosorbent assay for the measurements of CEA and PSA
Journal Title: Journal of Analytical Bio-Science - Year 2012, Vol 35, Issue 3
Abstract
To investigate the effects of supermolecules on the enzyme-linked immunosorbent assay (ELISA), different types of supermolecules (dendrimers, cyclodextrins, crown ethers, surfactants, and polyethylene glycols), and solid-phase sandwich ELISA kits for measurement of carcinoembryonic antigen (CEA) and prostate specific antigen (PSA) were used in this study. In the one-step CEA ELISA, each supermolecule was added to the capture antibody immobilized well. Standard and antibody-enzyme conjugates were then dispensed into the well. The subsequent procedures were performed as described in the user manual. In the two-step PSA ELISA, each supermolecule was added to the well either before dispensation of a standard and assay buffer in the first reaction, or before dispensation of an antibody-enzyme conjugate in the second reaction. Except for above mentioned process procedures, were performed as described in the user manual. In all experiments, the absorbance of color development by adding an enzymatic substrate in each well was measured and compared with that of a control prepared with saline instead of supermolecules. In the CEA measurement, absorbance was enhanced by the addition of Brij 35, Tween 20, PEG 6000, or PEG 4000. In the PSA measurement, absorbance was enhanced by the addition of PEG 6000 or PEG 4000 at the second reaction, whereas useful effects were not observed by the addition of supermolecules at the first reaction. Our results suggest that those supermolecules contributed to the enhancement of binding properties of antigen and enzyme-labeled antibody.
Authors and Affiliations
Tomoko Arai, Toshihiko Tsukada
Keywords
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