A comparative evaluation of microarray slides as substrates for the development of protease assay biosensors

作者：Darr Charles M; Castorena Jorge; Korampally Venumadhav; Gangopadhyay Shubhra^*; Polo Parada Luis

来源：Experimental and Molecular Pathology, 2011, 91(3): 714-717.

DOI：10.1016/j.yexmp.2011.09.009

• Microarray
• Botulinum neurotoxin A
• Protease biosensor

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 (R) 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 degrees C and a sub-lethal dose was detected in 2 hours.

• 出版日期2011-12