Determination of the Dissociation Constant for Polyvalent Receptors Using ELISA: A Case of M13 Phages Displaying Troponin T-Specific Peptides

Abstract

Phage-derived affinity peptides have become widespread thanks to their easy selection via phage display. Interactions between a target protein and its specific peptide are similar to those between antibodies and antigens. The strength of these noncovalent complexes may be described by the dissociation constant (K d ). In this paper, protein-specific peptides are exposed on the pIII protein present in the M13 bacteriophage virion with up to five copies. Therefore, one phage particle can bind from one to five ligands. Here, we discuss the dependences between phagedisplayed peptides and their ligands in solution using a model system based on troponin T (TnT) binding phages. Moreover, a method of calculating K d values from ELISA experiments was developed and is presented. The determined K d values are in the picomolar range.