Back to Search Start Over

Determining the Affinity and Kinetics of Small Molecule Inhibitors of Galectin-1 Using Surface Plasmon Resonance

Authors :
Henry Kim
Louis Kretz
Céline Ronin
Christina Starck
James A. Roper
Barbro Kahl-Knutson
Kristoffer Peterson
Hakon Leffler
Ulf J. Nilsson
Anders Pedersen
Fredrik R. Zetterberg
Robert J. Slack
Source :
International Journal of Molecular Sciences, Vol 25, Iss 12, p 6704 (2024)
Publication Year :
2024
Publisher :
MDPI AG, 2024.

Abstract

The beta-galactoside-binding mammalian lectin galectin-1 can bind, via its carbohydrate recognition domain (CRD), to various cell surface glycoproteins and has been implicated in a range of cancers. As a consequence of binding to sugar residues on cell surface receptors, it has been shown to have a pleiotropic effect across many cell types and mechanisms, resulting in immune system modulation and cancer progression. As a result, it has started to become a therapeutic target for both small and large molecules. In previous studies, we used fluorescence polarization (FP) assays to determine KD values to screen and triage small molecule glycomimetics that bind to the galectin-1 CRD. In this study, surface plasmon resonance (SPR) was used to compare human and mouse galectin-1 affinity measures with FP, as SPR has not been applied for compound screening against this galectin. Binding affinities for a selection of mono- and di-saccharides covering a 1000-fold range correlated well between FP and SPR assay formats for both human and mouse galectin-1. It was shown that slower dissociation drove the increased affinity at human galectin-1, whilst faster association was responsible for the effects in mouse galectin-1. This study demonstrates that SPR is a sound alternative to FP for early drug discovery screening and determining affinity estimates. Consequently, it also allows association and dissociation constants to be measured in a high-throughput manner for small molecule galectin-1 inhibitors.

Details

Language :
English
ISSN :
14220067 and 16616596
Volume :
25
Issue :
12
Database :
Directory of Open Access Journals
Journal :
International Journal of Molecular Sciences
Publication Type :
Academic Journal
Accession number :
edsdoj.401dc04e96c343c9aa7b54b26cf21dd8
Document Type :
article
Full Text :
https://doi.org/10.3390/ijms25126704