Journal
ANALYTICAL CHEMISTRY
Volume 84, Issue 4, Pages 1941-1947Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac300070t
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Funding
- National Science Foundation [DBI-0964216, DBI-0964137]
- NIH [1R01GM095638, R01 GM 092993]
- Office of Naval Research (ONR)
- NSF through the National Nanotechnology Infrastructure Network
- Direct For Biological Sciences
- Div Of Biological Infrastructure [1054191] Funding Source: National Science Foundation
- Direct For Biological Sciences
- Div Of Biological Infrastructure [0964216, 0964137] Funding Source: National Science Foundation
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We demonstrate an affordable low-noise surface plasmon resonance (SPR) instrument based on extraordinary optical transmission (EOT) in metallic nanohole arrays and quantify a broad range of antibody-ligand binding kinetics with equilibrium dissociation constants ranging from 200 pM to 40 nM. This nanohole-based SPR instrument is straightforward to construct, align, and operate, since it is built around a standard microscope and a portable fiber-optic spectrometer. The measured refractive index resolution of this platform is 3.1 x 10(-6) without on-chip cooling, which is among the lowest reported for SPR sensors based on EOT. This is accomplished via rapid full-spectrum acquisition in 10 ms followed by frame averaging of the EOT spectra, which is made possible by the production of template-stripped gold nanohole arrays with homogeneous optical properties over centimeter-sized areas. Sequential SPR measurements are performed using a 12-channel microfluidic flow cell after optimizing surface modification protocols and antibody injection conditions to minimize mass-transport artifacts. The immobilization of a model ligand, the protective antigen of anthrax on the gold surface, is monitored in real-time with a signal-to-noise ratio of similar to 860. Subsequently, real-time binding kinetic curves were measured quantitatively between the antigen and a panel of small, 25 kDa single-chain antibodies at concentrations down to 1 nM. These results indicate that nanohole-based SPR instruments have potential for quantitative antibody screening and as a general-purpose platform for integrating SPR sensors with other bioanalytical tools.
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