Reduction of nonspecific protein binding on surface plasmon resonance biosensors

Jean Francois Masson, Tina M. Battaglia, Jeff Cramer, Stephen Beaudoin, Michael Sierks, Karl S. Booksh

Research output: Contribution to journalArticle

56 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1951-1959
Number of pages9
JournalAnalytical and bioanalytical chemistry
Volume386
Issue number7-8
DOIs
StatePublished - Dec 1 2006

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Keywords

  • 11-Mercaptoundecanoic acid
  • 16-Mercaptohexadecanoic acid
  • Biomarker detection
  • CM dextran
  • Gold surfaces
  • Serum adsorption
  • Short-chain thiol

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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