Demonstration of non‐linear detection in ELISA resulting in up to 1000‐fold too high affinities of fibronogen binding to integrin αIIbβ3

FEBS Letters

Engel, Jürgen¹  Tangemann, Kirsten¹ 

[1] Biozentrum, University of Basel, Klingelbergstr. 70, CH-4056 Basel, Switzerland

关键词: Solid phase assay;    Detection;    Integrin αIIbβ3;    Fibrinogen;   

DOI  :  10.1016/0014-5793(94)01411-S

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To clarify the question as to why different solid-phase assays yield different results in terms of interaction strength, we used fibrinogen binding to immobilized αIIbβ3 integrin as a test system. A classical ‘three step’ enzyme-linked (ELISA), a ‘two step’ biotin enzyme-linked streptavidin and a ‘one step’ radioligand assay were compared under otherwise identical conditions. Only the last assay yielded binding constants comparable to earlier data by total internal reflection fluorescence microscopy while the other assays yielded apparent binding constants 5- to 1000-fold too high. These effects are explained by non-linearity of detection signals.

【 授权许可】

Unknown