Reduction of nonspecific protein binding on surface plasmon resonance biosensors

Reduction of the nonspecific serum protein adsorption on a gold surface to levels low enough to allow the detection of biomarkers in complex media has been achieved using the N-hydroxysuccinimide (NHS) ester of 16-mercaptohexadecanoic acid. Carboxymethylated dextran (CM dextran), which is widely use...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry 2006-12, Vol.386 (7-8), p.1951-1959
Main Authors: Masson, Jean-Francois, Battaglia, Tina M, Cramer, Jeff, Beaudoin, Stephen, Sierks, Michael, Booksh, Karl S
Format: Article
Language:English
Subjects:
adsorption
Animals
binding properties
biomarkers
biosensors
blood proteins
Blood Proteins - analysis
Blood Proteins - metabolism
blood serum
Cattle
cell culture
Culture Media
dextran
gold
Humans
interleukin-6
Interleukin-6 - analysis
Interleukin-6 - metabolism
myoglobin
Myoglobin - analysis
Myoglobin - metabolism
Protein Binding
Sensitivity and Specificity
surface plasmon resonance
Surface Plasmon Resonance - methods
thiols
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Summary:Reduction of the nonspecific serum protein adsorption on a gold surface to levels low enough to allow the detection of biomarkers in complex media has been achieved using the N-hydroxysuccinimide (NHS) ester of 16-mercaptohexadecanoic acid. Carboxymethylated dextran (CM dextran), which is widely used, nonspecifically adsorbs enough proteins to mask the signal from target biomarkers in complex solutions such as serum or blood. The use of short-chain thiols greatly reduces the amount of nonspecific protein adsorption. Mixed layers of 11-mercaptoundecanoic acid or the NHS ester of 11-mercaptoundecanoic acid mixed layers with either 11-mercaptoundecanol or undecanethiol, and 16-mercaptohexadecanoic acid or the NHS ester of 16-mercaptohexadecanoic acid with hexadecanethiol, were also investigated for nonspecific protein binding properties as well as for biomarker signal response. The NHS ester of 16-mercaptohexadecanoic acid exhibits the largest signal for the biomarker myoglobin (including CM dextran) while offering a significantly diminished amount of nonspecific binding. The sensor has also been shown to detect interleukin-6 in cell culture media containing protein concentrations of at least 4 mg/mL.
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-006-0834-2