Labelling of colloidal gold with protein A. A quantitative study.
Colloidal gold complexes with protein A are extensively used in immunocytochemistry as secondary reagents for the localization of antigens. However detailed information on the process and extent of adsorption of protein A onto gold particles, the optimal condition of preparation and the stability of such complexes are lacking. The adsorption isotherm of 125I-protein A onto gold particles (11.2 nm in diameter) was studied quantitatively with gold sols buffered at pH 4-7. At low coverage of the particles, the isotherm was independent of pH. However in the presence of a large excess of protein A, the highest coverage was obtained with a gold sol buffered at pH 5.1, the isoelectric point of the protein. The association constant was decreased at high coverage of the particles. Maximum binding of the complex to immobilized IgG occurred with particles labelled with at least 9 molecules of protein A. The complex was stable under storage with up to 12 molecules adsorbed per particle. At high coverage (26 molecules per particle), a progressive loss of protein A was observed. The optimum condition for preparing the complex are reported.
D000327:Adsorption; D003102:Colloids; D006046:Gold; D013205:Staphylococcal Protein A
D003102:Colloids; D013205:Staphylococcal Protein A; D006046:Gold