Abstract

Localized surface plasmon resonance (LSPR) spectroscopy and imaging are emerging biosensor technologies which tout label-free biomolecule detection at the nanoscale and ease of integration with standard microscopy setups. The applicability of these techniques can be limited by the restrictions that surface-conjugated ligands must be both sufficiently small and orientated to meet analyte sensitivity requirements. We demonstrate that orientated single domain antibodies (sdAb) can optimize nanoplasmonic sensitivity by comparing three anti-ricin sdAb constructs to biotin-neutravidin, a model system for small and highly orientated ligand studies. LSPR imaging of electrostatically orientated sdAb exhibited a ricin sensitivity equivalent to that of the biotinylated LSPR biosensors for neutravidin. These results, combined with the facts that sdAb are highly stable and readily produced in bacteria and yeast, build a compelling case for the increased utilization of sdAbs in nanoplasmonic applications.Electronic supplementary materialThe online version of this article (doi:10.1007/s11468-015-9969-3) contains supplementary material, which is available to authorized users.

Highlights

  • Localized surface plasmon resonance (LSPR) biosensing is a label-free technique that enables biomolecule detection with nanoscale spatial resolution

  • Using SPR imaging (SPRi), we show that these modified single domain antibodies (sdAb) have enhanced surface conjugation efficiencies and improved sensitivity to ricin relative to the non-orientated sdAb

  • Three ligands were used in this study: an unmodified sdAb (C8), a genetic fusion of C8 with a positively charged peptide (C8-zip), and a genetic fusion of the D12f sdAb with the biotin binding protein rhizavidin (D12f-rhiz)

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Summary

Introduction

Localized surface plasmon resonance (LSPR) biosensing is a label-free technique that enables biomolecule detection with nanoscale spatial resolution. ScFvs and Fab fragments with molecular weights of approximately 27 and 50 kDa, respectively, are still relatively large ligands for many LSPR biosensing applications, leaving room for improved sensitivity if the size can be further reduced.

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