Orientation of llama antibodies strongly increases sensitivity of biosensors

A.K. Trilling, T. Hesselink, A. van Houwelingen, J.H.G. Cordewener, M.A. Jongsma, S. Schoffelen, J.C.M. van Hest, J.T. Zuilhof, J. Beekwilder

Research output: Contribution to journalArticleAcademicpeer-review

24 Citations (Scopus)

Abstract

Sensitivity of biosensors depends on theorientation of bio-receptors on the sensor surface.The objective of this study was to organize bio-receptors on surfaces in a way that their analyte binding site is exposed to the analyte solution. VHH proteins recognizing foot-and-mouth disease virus (FMDV) were used for making biosensors, and azides were introduced in the VHH to function as bioorthogonal reactive groups. The importance of the orientation of bio-receptors was addressed by comparing sensors with randomly oriented VHH (with multiple exposed azide groups)to sensors with uniformly oriented VHH (with only a single azide group). A surface plasmon resonance (SPR) chip exposing cyclooctyne was reacted to azide functionalized VHH domains, using click chemistry. Comparison between randomly and uniformly oriented bio-receptors showed up to 800-fold increase in biosensor sensitivity. This technique may increase the containment of infectious diseases such as FMDV as its strongly enhanced sensitivity may facilitate early diagnostics.
LanguageEnglish
Pages130-136
JournalBiosensors and Bioelectronics
Volume60
DOIs
Publication statusPublished - 2014

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New World Camelids
Azides
Biosensing Techniques
Biosensors
Antibodies
Foot-and-Mouth Disease Virus
Viruses
Sensors
Click Chemistry
Surface Plasmon Resonance
Surface plasmon resonance
Binding sites
Communicable Diseases
Binding Sites
Proteins

Keywords

  • autodisplayed z-domains
  • click chemistry
  • immobilization
  • coli

Cite this

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title = "Orientation of llama antibodies strongly increases sensitivity of biosensors",
abstract = "Sensitivity of biosensors depends on theorientation of bio-receptors on the sensor surface.The objective of this study was to organize bio-receptors on surfaces in a way that their analyte binding site is exposed to the analyte solution. VHH proteins recognizing foot-and-mouth disease virus (FMDV) were used for making biosensors, and azides were introduced in the VHH to function as bioorthogonal reactive groups. The importance of the orientation of bio-receptors was addressed by comparing sensors with randomly oriented VHH (with multiple exposed azide groups)to sensors with uniformly oriented VHH (with only a single azide group). A surface plasmon resonance (SPR) chip exposing cyclooctyne was reacted to azide functionalized VHH domains, using click chemistry. Comparison between randomly and uniformly oriented bio-receptors showed up to 800-fold increase in biosensor sensitivity. This technique may increase the containment of infectious diseases such as FMDV as its strongly enhanced sensitivity may facilitate early diagnostics.",
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year = "2014",
doi = "10.1016/j.bios.2014.04.017",
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Orientation of llama antibodies strongly increases sensitivity of biosensors. / Trilling, A.K.; Hesselink, T.; van Houwelingen, A.; Cordewener, J.H.G.; Jongsma, M.A.; Schoffelen, S.; van Hest, J.C.M.; Zuilhof, J.T.; Beekwilder, J.

In: Biosensors and Bioelectronics, Vol. 60, 2014, p. 130-136.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Orientation of llama antibodies strongly increases sensitivity of biosensors

AU - Trilling, A.K.

AU - Hesselink, T.

AU - van Houwelingen, A.

AU - Cordewener, J.H.G.

AU - Jongsma, M.A.

AU - Schoffelen, S.

AU - van Hest, J.C.M.

AU - Zuilhof, J.T.

AU - Beekwilder, J.

PY - 2014

Y1 - 2014

N2 - Sensitivity of biosensors depends on theorientation of bio-receptors on the sensor surface.The objective of this study was to organize bio-receptors on surfaces in a way that their analyte binding site is exposed to the analyte solution. VHH proteins recognizing foot-and-mouth disease virus (FMDV) were used for making biosensors, and azides were introduced in the VHH to function as bioorthogonal reactive groups. The importance of the orientation of bio-receptors was addressed by comparing sensors with randomly oriented VHH (with multiple exposed azide groups)to sensors with uniformly oriented VHH (with only a single azide group). A surface plasmon resonance (SPR) chip exposing cyclooctyne was reacted to azide functionalized VHH domains, using click chemistry. Comparison between randomly and uniformly oriented bio-receptors showed up to 800-fold increase in biosensor sensitivity. This technique may increase the containment of infectious diseases such as FMDV as its strongly enhanced sensitivity may facilitate early diagnostics.

AB - Sensitivity of biosensors depends on theorientation of bio-receptors on the sensor surface.The objective of this study was to organize bio-receptors on surfaces in a way that their analyte binding site is exposed to the analyte solution. VHH proteins recognizing foot-and-mouth disease virus (FMDV) were used for making biosensors, and azides were introduced in the VHH to function as bioorthogonal reactive groups. The importance of the orientation of bio-receptors was addressed by comparing sensors with randomly oriented VHH (with multiple exposed azide groups)to sensors with uniformly oriented VHH (with only a single azide group). A surface plasmon resonance (SPR) chip exposing cyclooctyne was reacted to azide functionalized VHH domains, using click chemistry. Comparison between randomly and uniformly oriented bio-receptors showed up to 800-fold increase in biosensor sensitivity. This technique may increase the containment of infectious diseases such as FMDV as its strongly enhanced sensitivity may facilitate early diagnostics.

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