Measurements of Protein-Protein Interactions by Size Exclusion Chromatography

TL;DR

This paper introduces a size exclusion chromatography method to measure protein-protein interactions via second virial coefficients, aligning well with light scattering data and tracking interaction changes.

Contribution

It presents a novel SEC-based approach for determining second virial coefficients of proteins, enabling analysis of interaction evolution in solutions.

Findings

SEC method accurately measures B_2 values

Method tracks transition from positive to negative B_2

Results agree with light scattering data

Abstract

A method is presented for determining second virial coefficients B_2 of protein solutions from retention time measurements in size exclusion chromatography (SEC). We determine B_2 by analyzing the concentration dependance of the chromatographic partition coefficient. We show the ability of this method to track the evolution of B_2 from positive to negative values in lysozyme and bovine serum albumin solutions. Our SEC results agree quantitatively with data obtained by light scattering.