Abstract

Non-specific adsorption (NSA) of biomolecules is a persistent challenge in microfluidic biosensors. Microfluidic biosensors often have immobilized bioreceptors such as antibodies, enzymes, DNAs, etc, via linker molecules such as SAMs (self-assembled monolayers) to enhance immobilization. However, the linker molecules are very susceptible to NSA, causing false responses and decreasing sensitivity. In this paper, we present design methods to reduce the NSA of alkanethiol SAMs, which are popular linker molecules on microfluidic biosensors. Three design parameters were studied for two different chain-length SAMs (n = 2 and 10): (i) SAM incubation time, (ii) surface roughness [0.8 nm and 4.4 nm RMS (root mean square)] and (iii) gold crystal re-growth along (1 1 1) the target orientation. NSA was monitored by surface plasmon resonance (SPR). The results suggest that increased SAM incubation time reduces NSA, and that short-chain SAMs respond more favorably than the long-chain SAMs. Both SAMs were shown to be sensitive to surface roughness, and long-chain SAMs reduced NSA by 75%. Gold crystal re-growth along (1 1 1) the target orientation profoundly reduced NSA on the short-chain SAM. On a gold surface where surface roughness was 0.8 nm and there was strong directional alignment along the (1 1 1) gold crystal, final concentrations of nonspecifically bound proteins were 0.05 ng mm-2 (fibrinogen) and 0.075 ng mm-2 (lysozyme)- significantly lower than other known methods. The results show that optimizing three parameters (SAM incubation time, gold surface roughness and gold crystal orientation) improved SAM sensitivity for fibrinogen-anti-fibrinogen conjugates by a factor of 5 in 2.94 pM, suggesting that the methods are effective for reducing NSA in microfluidic biosensors.

Original languageEnglish (US)
Article number075015
JournalJournal of Micromechanics and Microengineering
Volume20
Issue number7
DOIs
StatePublished - 2010

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Self assembled monolayers
Microfluidics
Biosensors
Adsorption
Gold
Surface roughness
Fibrinogen
Crystals
Molecules
Enzymes
Surface plasmon resonance
Biomolecules
Muramidase
Chain length
Antibodies
Crystal orientation
DNA
Proteins

ASJC Scopus subject areas

  • Mechanical Engineering
  • Electrical and Electronic Engineering
  • Mechanics of Materials
  • Electronic, Optical and Magnetic Materials

Cite this

Methods of reducing non-specific adsorption in microfluidic biosensors. / Choi, Seokheun; Chae, Junseok.

In: Journal of Micromechanics and Microengineering, Vol. 20, No. 7, 075015, 2010.

Research output: Contribution to journalArticle

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abstract = "Non-specific adsorption (NSA) of biomolecules is a persistent challenge in microfluidic biosensors. Microfluidic biosensors often have immobilized bioreceptors such as antibodies, enzymes, DNAs, etc, via linker molecules such as SAMs (self-assembled monolayers) to enhance immobilization. However, the linker molecules are very susceptible to NSA, causing false responses and decreasing sensitivity. In this paper, we present design methods to reduce the NSA of alkanethiol SAMs, which are popular linker molecules on microfluidic biosensors. Three design parameters were studied for two different chain-length SAMs (n = 2 and 10): (i) SAM incubation time, (ii) surface roughness [0.8 nm and 4.4 nm RMS (root mean square)] and (iii) gold crystal re-growth along (1 1 1) the target orientation. NSA was monitored by surface plasmon resonance (SPR). The results suggest that increased SAM incubation time reduces NSA, and that short-chain SAMs respond more favorably than the long-chain SAMs. Both SAMs were shown to be sensitive to surface roughness, and long-chain SAMs reduced NSA by 75{\%}. Gold crystal re-growth along (1 1 1) the target orientation profoundly reduced NSA on the short-chain SAM. On a gold surface where surface roughness was 0.8 nm and there was strong directional alignment along the (1 1 1) gold crystal, final concentrations of nonspecifically bound proteins were 0.05 ng mm-2 (fibrinogen) and 0.075 ng mm-2 (lysozyme)- significantly lower than other known methods. The results show that optimizing three parameters (SAM incubation time, gold surface roughness and gold crystal orientation) improved SAM sensitivity for fibrinogen-anti-fibrinogen conjugates by a factor of 5 in 2.94 pM, suggesting that the methods are effective for reducing NSA in microfluidic biosensors.",
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