Issue 8, 2023

Sortase A transpeptidation produces seamless, unbranched biotinylated nanobodies for multivalent and multifunctional applications

Abstract

Exploitation of the biotin–streptavidin interaction for advanced protein engineering is used in many bio-nanotechnology applications. As such, researchers have used diverse techniques involving chemical and enzyme reactions to conjugate biotin to biomolecules of interest for subsequent docking onto streptavidin-associated molecules. Unfortunately, the biotin–streptavidin interaction is susceptible to steric hindrance and conformational malformation, leading to random orientations that ultimately impair the function of the displayed biomolecule. To minimize steric conflicts, we employ sortase A transpeptidation to produce quantitative, seamless, and unbranched nanobody–biotin conjugates for efficient display on streptavidin-associated nanoparticles. We further characterize the protein–nanoparticle complex and demonstrate its usefulness in optical microscopy and multivalent severe acute respiratory syndrome coronavirus (SARS-CoV-2) antigen interaction. The approach reported here provides a template for making novel multivalent and multifunctional protein complexes for avidity-inspired technologies.

Graphical abstract: Sortase A transpeptidation produces seamless, unbranched biotinylated nanobodies for multivalent and multifunctional applications

Supplementary files

Article information

Article type
Paper
Submitted
06 Jan 2023
Accepted
15 Mar 2023
First published
15 Mar 2023
This article is Open Access
Creative Commons BY-NC license

Nanoscale Adv., 2023,5, 2251-2260

Sortase A transpeptidation produces seamless, unbranched biotinylated nanobodies for multivalent and multifunctional applications

E. M. Obeng, D. L. Steer, A. J. Fulcher and K. M. Wagstaff, Nanoscale Adv., 2023, 5, 2251 DOI: 10.1039/D3NA00014A

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