This application note investigates the effect of NaCl concentration on the elution volume and separation of proteins by size exclusion chromatography.
Size exclusion chromatography (SEC) is a technique used to determine the molecular size of proteins and polymers. The presence or absence of salts, such as sodium chloride (NaCl), in the mobile phase can influence protein elution volumes by altering their hydrodynamic radius and secondary interactions with the resin.
This document investigates the retention behavior of three globular proteins—y-globulins, bovine serum albumin, and ovalbumin—using an Agilent ProSEC 3008 column. The study compares UV responses at varying NaCl concentrations to determine how salt strength affects non-ideal protein-protein and protein-matrix interactions.
The purpose is to minimize secondary interactions of the sample components with the resin.
It alters their hydrodynamic radius as a function of salt strength.
The column dimensions are 300 x 7.5 mm.