biostudies-literatureNeves MMPSEuropean Research CouncilUniversity of WarwickBiotechnology and Biological Sciences Research CouncilH2020 Marie Skłodowska-Curie ActionsEngineering and Physical Sciences Research Council377-382https://www.ebi.ac.uk/biostudies/studies/S-EPMC9969229biostudies-literatureUnknown8(3)Glycoforms (and other post-translational modifications) of otherwise identical proteins can indicate pathogenesis/disease state and hence new tools to detect and sense a protein's glycosylation status are essential. Antibody-based assays against specific protein sequences do not typically discriminate between glycoforms. Here we demonstrate a 'sandwich' bio-assay approach, whereby antibodies immobilised onto biolayer interferometry sensors first select proteins, and then the specific glycoform is identified using gold nanoparticles functionalised with lectins which provide signal enhancement. The nanoparticles significantly enhance the signal relative to lectins alone, allowing glycoform specific detection as low as 0.04 μg mL^-1 (1.4 nM) in buffer, and crucially there is no needNanoscale horizonsDiscrimination between protein glycoforms using lectin-functionalised gold nanoparticles as signal enhancers.PMC9969229EP/R511808/1BB/S506783/1713548866056Baker ANNeves MMPSWalker MRichards SJGeorgiou PGGibson MIfalseDiscrimination between protein glycoforms using lectin-functionalised gold nanoparticles as signal enhancers.Glycoforms (and other post-translational modifications) of otherwise identical proteins can indicate pathogenesis/disease state and hence new tools to detect and sense a protein's glycosylation status are essential. Antibody-based assays against specific protein sequences do not typically discriminate between glycoforms. Here we demonstrate a 'sandwich' bio-assay approach, whereby antibodies immobilised onto biolayer interferometry sensors first select proteins, and then the specific glycoform is identified using gold nanoparticles functionalised with lectins which provide signal enhancement. The nanoparticles significantly enhance the signal relative to lectins alone, allowing glycoform specific detection as low as 0.04 μg mL^-1 (1.4 nM) in buffer, and crucially there is no need2023-01-01T00:00:00Z2023 Feb2026-03-17T15:57:40.047Z2025-04-04T21:48:19.968ZS-EPMC99692293665129210.1039/d2nh00470d