Cell-free expression and characterization of multivalent rhamnose-binding lectins using biolayer interferometry

{Lectins are important biological tools for binding glycans, but recombinant protein expression poses challenges for some lectin classes, limiting the pace of discovery and characterization. To discover and engineer lectins with new functions, workflows amenable to rapid expression and subsequent characterization are needed. Here, we present bacterial cell-free protein synthesis as a means for efficient, small-scale expression of multivalent, disulfide bond-rich, rhamnose-binding lectins. Furthermore, we demonstrate that the cell-free expressed lectins can be directly coupled with bio-layer interferometry (BLI) analysis, either in solution or immobilized on the sensor, to measure interaction with carbohydrate ligands without purification. This workflow enables determination of lectin substrate specificity and estimation of binding affinity. Overall, we believe that this method will enable high-throughput expression, screening, and characterization of new and engineered multivalent lectins for applications in synthetic glycobiology.}

Références

Titre
Cell-free expression and characterization of multivalent rhamnose-binding lectins using biolayer interferometry
Type de publication
Article de revue
Année de publication
2023
Revue
Glycobiology
Date de publication
03/2023
ISSN
1460-2423
Soumis le 20 mars 2023