Kricka L J, Carter T J, Burt S M, Kennedy J H, Holder R L, Halliday M I, Telford M E, Wisdom G B
Clin Chem. 1980 May;26(6):741-4.
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.
我们通过使用涉及酶 - 蛋白质缀合物的程序，评估了吸附在各种微量滴定板单个孔表面的蛋白质量的变异性。吸附蛋白的变异系数范围为5.2%至29.5%。微量滴定板显示出明显的“边缘效应”；板边缘的孔比内部的孔吸附更多的蛋白质，这显著影响了涉及此类板的免疫测定结果。在夹心酶免疫测定中，单个样品的结果变化为±18%；在竞争测定中，同一板上不同校准曲线读取的值变化为±11%。扫描电子显微镜和双折射研究表明，板上单个孔之间没有明显的物理差异。吸附在同一微量滴定板单个孔表面的蛋白质量的变异性严重影响了将此类板用作固相支持物的免疫测定结果的可靠性和解读。
