Label-free reading of microarray-based immunoassays with surface plasmon resonance imaging.
A simple method is presented for patterning of protein antigens at a gold surface for use in surface plasmon resonance (SPR) imaging experiments. Microfluidic devices fabricated from poly(dimethylsiloxane) were used to flow various fluids over a gold substrate in spatially defined channels. This technique was used to pattern the surface chemistry of the gold as well as to adsorb antigens from solution to the modified substrates. The resulting antigen arrays were probed with complementary antibodies in order to demonstrate the effectiveness of the patterning for antibody capture experiments. SPR imaging was used to aid in the optimization of array fabrication and to observe the interactions of unlabeled antibodies with these microarrays. This work presents a means of fabricating microarrays with controlled surface density of antigens. SPR imaging provides both quantitative and qualitative evaluation of antibody binding in a label free format.
Kanda
Vishal
V
Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2 Canada.
Kariuki
James K
JK
Harrison
D Jed
DJ
McDermott
Mark T
MT
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000941:Antigens / Q000737:chemistry; D002417:Cattle; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D044085:Microfluidics / Q000379:methods; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*
D000906:Antibodies; D000941:Antigens; D004129:Dimethylpolysiloxanes; D007074:Immunoglobulin G