Covalent immobilization of proteins onto photoactivated polystyrene microtiter plates for enzyme-linked immunosorbent assay procedures.
Enhancement of the speed and sensitivity of an ELISA technique was achieved by doing it on a polystyrene microtiter plate preactivated by a simple photochemical reaction. Immobilization of Epicoccum nigrum antigen (allergenic antigen) or goat anti-rabbit IgG onto the photoactivated plates was found to occur in only 45 min with higher binding than that obtained through adsorption during the same period onto the untreated surface. Nearly 1.5-2-folds higher readings were obtained when the ELISA was carried out with the solid phase prepared on the photoactivated surface rather than on the untreated surface. Moreover, solid phases prepared on the activated surface could detect IgE (E. nigrum antibody) even at 1/50 (v/v) dilutions, whereas a solid phase prepared on the untreated surface failed to do so. Around three times higher ELISA values were obtained in the activated plate than the untreated plate when IgE was diluted to 1/5 (v/v). Such photoactivated surface could be of great importance in diagnostic tests involving the ELISA technique particularly to confirm false negative cases and for other immunoassays such as radioimmunoassay procedures.
Bora
Utpal
U
Centre for Biochemical Technology (CSIR), Delhi University Campus, Mall Road, India.
Chugh
Lalita
L
Nahar
Pradip
P
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006041:Goats; D007073:Immunoglobulin E / Q000032:analysis; D007074:Immunoglobulin G / Q000276:immunology; D010777:Photochemistry; D011137:Polystyrenes; D011817:Rabbits
D007074:Immunoglobulin G; D011137:Polystyrenes; D007073:Immunoglobulin E