title: A comparative evaluation of microarray slides as substrates for the development of protease assay biosensors.
abstract: The application of commercially available microarray slides as substrates for fluorogenic protease assays has been explored in terms of binding efficiency, stability, and activity. A fluorescent, biotinylated substrate for botulinum neurotoxin A (BoNTA) was attached via self-assembled monolayer of Streptavidin to amine-reactive aldehyde, epoxy, hydrogel, and polymer slides. Nexterion Slide P® was found to have optimal protein binding efficiency and stability of the slides examined. Addition of glycerol to the printing buffer improved spot morphology significantly and polyvinylpyrrolidone provided long-term stability, allowing chips to be stored for up to 1 month with good viability. Detection of a recombinant BoNTA light chain was then carried out at 37°C and a sub-lethal dose was detected in 2 hours.
authors.affiliation: Department of Biological Engineering, University of Missouri-Columbia, Columbia, MO, USA.
authors.forename: Charles M
authors.initials: CM
authors.lastname: Darr
authors.forename: Jorge
authors.initials: J
authors.lastname: Castorena
authors.forename: Venumadhav
authors.initials: V
authors.lastname: Korampally
authors.forename: Shubhra
authors.initials: S
authors.lastname: Gangopadhyay
authors.forename: Luis
authors.initials: L
authors.lastname: Polo-Parada
mesh_terms: D000447:Aldehydes; D001681:Biological Assay / Q000295:instrumentation; D015374:Biosensing Techniques; D002021:Buffers; D004852:Epoxy Compounds; D005990:Glycerol; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D010447:Peptide Hydrolases / Q000032:analysis*; D011108:Polymers; D019809:Streptavidin
chemical_list: D000447:Aldehydes; D002021:Buffers; D004852:Epoxy Compounds; D011108:Polymers; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D019809:Streptavidin; D010447:Peptide Hydrolases; D005990:Glycerol