Glycoforms (and different post-translational modifications) of in any other case equivalent proteins can point out pathogenesis/illness state and therefore new instruments to detect and sense a protein’s glycosylation standing are important. Antibody-based assays towards particular protein sequences don’t usually discriminate between glycoforms. Right here we show a ‘sandwich’ bio-assay method, whereby antibodies immobilised onto biolayer interferometry sensors first choose proteins, after which the particular glycoform is recognized utilizing gold nanoparticles functionalised with lectins which offer sign enhancement. The nanoparticles considerably improve the sign relative to lectins alone, permitting glycoform particular detection as little as 0.04 μg mL−1 (1.4 nM) in buffer, and crucially there is no such thing as a want for an enrichment step and all steps may be automated. Proof of idea is demonstrated utilizing prostate particular antigen: a biomarker for prostate most cancers, the place glycoform evaluation might distinguish between cancerous and non-cancerous standing, somewhat than solely detecting general protein focus.

Graphical abstract: Discrimination between protein glycoforms using lectin-functionalised gold nanoparticles as signal enhancers

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