Clinical Chemistry, Vol 26, 741-744, Copyright © 1980 by American Association for Clinical Chemistry

Variability in the adsorption properties of microtitre plates used as solid supports in enzyme immunoassay

LJ Kricka, TJ Carter, SM Burt, JH Kennedy, RL Holder, MI Halliday, ME Telford and GB Wisdom

We evaluated the variability in the amount of protein adsorbed onto the surface of individual wells of an assortment of microtitre plates by use of procedures involving enzyme-protein conjugates. Coefficients of variation in adsorbed protein ranged from 5.2 to 29.5%. Microtitre plates show a distinct "edge effect"; wells at the edges of a plate adsorb more protein than those in the interior, which markedly affects results from immunoassays involving such plates. In a sandwich enzyme immunoassay the results for an individual sample varied by +/- 18%; in a competitive assay, values read from different calibration curves in the same plate varied by +/- 11%. Scanning electron microscopy and birefringence studies demonstrated no marked physical differences between individual wells on a plate. The variability in the amount of protein adsorbed onto the surface of individual wells on the same microtitre plate seriously affects the reliability and interpretation of results of immunoassays in which such plates are used as solid supports.

The following articles in journals at HighWire Press have cited this article:

				L. J. Kricka and S. R. Master Quality Control and Protein Microarrays Clin. Chem., June 1, 2009; 55(6): 1053 - 1055. [Full Text] [PDF]

				B. K. Lundholt, K. M. Scudder, and L. Pagliaro A Simple Technique for Reducing Edge Effect in Cell-Based Assays J Biomol Screen, October 1, 2003; 8(5): 566 - 570. [Abstract] [PDF]