Quantitative Label-Free Characterization of Avidin-Biotin Assemblies on Silanized Glass

Title
Quantitative Label-Free Characterization of Avidin-Biotin Assemblies on Silanized Glass
Author(s)
서정현리정첸[리정첸]엘러[엘러]버크트로프[버크트로프]써니사하[써니사하]알렉산더레브진[알렉산더레브진]슈바이커츠[슈바이커츠]
Keywords
SECONDARY-ION EMISSION; MASS-SPECTROMETRY; TOF-SIMS; PROTEIN
Issue Date
201109
Publisher
AMER CHEMICAL SOC
Citation
ANALYTICAL CHEMISTRY, v.83, no.18, pp.7173 - 7178
Abstract
In this study, a time-of-flight secondary ion mass spectrometer TOF-SIMS, operating in the event-by-event bombardment/detection mode was used to characterize avidin-biotin assemblies on silane-modified glass substrates. SIMS was used to analyze several variants of the biointerface, including avidin physically adsorbed on a monofunctional acryl silane surface and covalently attached on monofunctional (amine terminated) and bifunctional (amine and acryl terminated) silanes. The goal of these studies was to determine density of avidin and biotin layers chemically or physically adsorbed on silanized glass substrate. An individual impact of a C(60) projectile used in this study creates a hemispherical crater (similar to 10 nm in diameter) and emits large numbers of secondary ions from the same nanovolume. Thus, a single impact enables one to unfold distinct secondary ions that span the thickness of the assembled film. This method was used to monitor the presence of glass, silane, and protein ions and to estimate the thickness and density of the avidin layer. In addition, we employed the double coincidence mass spectrometry approach to identify ions coemitted from a specific stratum of the biointerface. This approach was used to determine density of biotin and avidin immobilization while eliminating interferences from isobaric ions that originated from other constituents on the surface. Overall, novel TOF-SIMS quantitative approaches employed here were useful for examining complex biointerfaces and determining both lateral and in depth composition of the film.
URI
http://hdl.handle.net/YU.REPOSITORY/24641http://dx.doi.org/10.1021/ac2016085
ISSN
0003-2700
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공과대학 > 화학공학부 > Articles
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