Direct Immobilization of Engineered Nanobodies on Gold Sensors

Research output: Contribution to journalArticlepeer-review

Authors

  • Bárbara Simões
  • Wanda J Guedens
  • Charlie Keene
  • Karina Kubiak-Ossowska
  • Paul Mulheran
  • Anna M Kotowska
  • David J Scurr
  • Morgan R Alexander
  • Alexis Broisat
  • Steven Johnson
  • Serge Muyldermans
  • Nick Devoogdt
  • Peter Adriaensens

External organisations

  • Hasselt University
  • University of Strathclyde
  • University of Nottingham
  • Université Grenoble Alpes
  • York University
  • University of Liverpool
  • Vrije Universiteit Brussel

Abstract

Single-domain antibodies, known as nanobodies, have great potential as biorecognition elements for sensors because of their small size, affinity, specificity, and robustness. However, facile and efficient methods of nanobody immobilization are sought that retain their maximum functionality. Herein, we describe the direct immobilization of nanobodies on gold sensors by exploiting a modified cysteine strategically positioned at the C-terminal end of the nanobody. The experimental data based on secondary ion mass spectrometry, circular dichroism, and surface plasmon resonance, taken together with a detailed computational work (molecular dynamics simulations), support the formation of stable and well-oriented nanobody monolayers. Furthermore, the nanobody structure and activity is preserved, wherein the nanobody is immobilized at a high density (approximately 1 nanobody per 13 nm2). The strategy for the spontaneous nanobody self-assembly is simple and effective and possesses exceptional potential to be used in numerous sensing platforms, ranging from clinical diagnosis to environmental monitoring.

Details

Original languageEnglish
Pages (from-to)17353-17360
Number of pages8
JournalACS Applied Materials & Interfaces
Volume13
Issue number15
Early online date12 Apr 2021
Publication statusPublished - 21 Apr 2021