Hybrid nanoparticle-liposome detection of phospholipase activity.
A flexible nanoparticle-based phospholipase (PL) assay is demonstrated in which the enzymatic substrate is decoupled from the nanoparticle surface. Liposomes are loaded with a polypeptide that is designed to heteroassociate with a second polypeptide immobilized on gold nanoparticles. Release of this polypeptide from the liposomes, triggered by PL, induces a folding-dependent nanoparticle bridging aggregation. The colorimetric response from this aggregation enables straightforward and continuous detection of PL in the picomolar range. The speed, specificity, and flexibility of this assay make it appropriate for a range of applications, from point of care diagnostics to high-throughput pharmaceutical screening.
Aili
Daniel
D
Department of Materials and Institute of Biomedical Engineering, Imperial College London, SW7 2AZ London, UK.
Mager
Morgan
M
Roche
David
D
Stevens
Molly M
MM
D003124:Colorimetry / Q000379:methods*; D004789:Enzyme Activation; D008081:Liposomes / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D010740:Phospholipases / Q000032:analysis* / Q000737:chemistry*
D008081:Liposomes; D010740:Phospholipases