Microcontact printed antibodies on gold surfaces: function, uniformity, and silicone contamination.
The function of microcontact printed protein was investigated using surface plasmon resonance (SPR) imaging, X-ray photoelectron spectroscopy spectroscopy (XPS), and XPS imaging. We chose to analyze a model protein system, the binding of an antibody from solution to a microcontact printed protein antigen immobilized to a gold surface. SPR imaging experiments indicated that the microcontact printed protein antigen was less homogeneous, had increased nonspecific binding, and bound less antibody than substrates to which the protein antigen had been physically adsorbed. SPR images of substrates contacted with a poly(dimethylsiloxane) stamp inked with buffer alone (i.e., no protein) revealed that significant amounts of silicone oligomer were transferred to the surface. The transfer of the silicone oligomer was not homogeneous, and the oligomer nonspecifically bound protein (BSA and IgG) from solution. XPS spectroscopy and imaging were used to quantify the amount of silicon (due to the presence of silicone oligomer), as well as the amounts of other elements, transferred to the surface. The results suggest that the silicone oligomer introduced by the printing process reduces the overall binding capacity of the microcontact-printed protein compared to physically adsorbed protein.
Foley
Jennifer O
JO
Department of Bioengineering and National ESCA and Surface Analysis Center for Biomedical Problems, University of Washington, Seattle, Washington 98195, USA.
Fu
Elain
E
Gamble
Lara J
LJ
Yager
Paul
P
D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D004340:Drug Contamination; D006046:Gold / Q000737:chemistry*; D054802:Molecular Imprinting / Q000379:methods*; D011485:Protein Binding; D011506:Proteins; D012828:Silicones; D013057:Spectrum Analysis; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D014965:X-Rays
D000906:Antibodies; D000941:Antigens; D011506:Proteins; D012828:Silicones; D006046:Gold