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Updated: Apr 27, 2026

Identification of Protein Interacting Partners Using Tandem Affinity Purification
Published on: February 25, 2012
Affinity assays for decrypting protein targets of unknown function
Matthew J Todd1, Maxwell D Cummings1, Marina I Nelen1
1Johnson & Johnson Pharmaceutical Research & Development, L.L.C., 665 Stockton Drive, Exton, PA 19341, USA.
Abstract:
The application of label-independent biophysical assays (affinity assays) to carefully selected libraries of biochemicals, natural products and known bioactive molecules generates a powerful tool to identify ligands indicative of protein function. Detection of ligand binding does not require the measurement of enzyme activity, thus every cofactor, ion, reactant or product might be identified separately. One such assay, ThermoFluor(®), monitors ligand perturbation of protein thermostability. In conjunction with a functional probe library (FPL), this assay has been used to decrypt the function of orphan proteins.:
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