APPLICATION OF COLLOIDAL GOLD FOR CHARACTERIZATION OF SUPPORTS USED IN SIZE-EXCLUSION CHROMATOGRAPHY

Calibration of matrices for size-exclusion (gel permeation) chromatography (SEC) has to be done for the characterization of supports with inert pores and to allow the determination of the molecular masses of macromolecules. It has been shown by several groups that for steric reasons SEC is very sensitive to conformational details of macromolecules (e.g., rods or random coils). Therefore, no set of particles, organic macromolecules or biomacromolecules such as proteins exists which is universally applicable for calibration and characterization of chromatographic supports. It is shown that colloidal gold, which behaves in SEC as macromolecules, offers the advantage of hydrophilic particles with ideally spherical form and easily measured diameters. Therefore. collodial gold can be used for monitoring of the SEC process at the electron microscopic level, for the characterization of supports (determination of pore size) and for the determination of exclusion limits of matrices.