Multiplex protein interaction determinations using glutathione-GST binding

摘要

Fusion proteins in which glutathione S-transferase (GST) is a fusion partner are used as an immobilized binding member in screening procedures or other multi-analyte test procedures based on protein interaction. In such procedures, a particular protein is selected from a group of candidate proteins on the basis of the binding affinity of that protein for a target protein, with either the candidate proteins or the target protein being a fusion partner with GST and the GST portion of the fusion partner having been immobilized on glutathione-coated particles by the binding of GST to glutathione. The particles themselves are classifiable by different values of a differentiation parameter that permits them to be distinguished by flow cytometry, and the procedure is conducted in a manner that associates the individual candidate proteins with individual classes of the particles. When a binding interaction occurs between a candidate protein and the target protein, the particles on which the interaction has occurred are readily distinguished by flow cytometry and correlated with the candidate protein that exhibited the binding.