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

26 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.
Original languageEnglish
Pages (from-to)130-136
JournalBiosensors and Bioelectronics
Volume60
DOIs
Publication statusPublished - 2014

Keywords

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

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