We move beyond antibody-antigen binding systems and demonstrate that short peptide ligands can be used to efficiently capture Bacillus subtilis (a simulant of Bacillus anthracis) spores in liquids. On an eight-cantilever array chip, four cantilevers were coated with binding peptide (NHFLPKV-GGGC) and the other four were coated with control peptide (LFNKHVP-GGGC) for reagentless detection of whole B. subtilis spores in liquids. The peptide-ligand-functionalized microcantilever chip was mounted onto a fluid cell filled with a B. subtilis spore suspension for approximately 40 min; a 40 nm net differential deflection was observed. Fifth-mode resonant frequency measurements were also performed before and after dipping microcantilever arrays into a static B. subtilis solution showing a substantial decrease in frequency for binding-peptide-coated microcantilevers as compared to that for control peptide cantilevers. Further confirmation was obtained by subsequent examination of the microcantilever arrays under a dark-field microscope. Applications of this technology will serve as a platform for the detection of pathogenic organisms including biowarfare agents.
Microcantilevers are often deployed in flowing fluids to measure local flow velocities or to detect rapidly the nanomechanical binding of trace quantities of target analytes. The authors investigate the flow-induced mechanics of microcantilevers by deriving a semianalytical theoretical model for the nanoscale deflections of an elastic microcantilever due to a laminar viscous flow incident upon it. Conversely, the model allows for the estimation of the local flow velocities based on measured microcantilever deflection. Careful experiments performed on silicon microcantilevers in flowing nitrogen confirm the theoretical predictions up to a critical flow rate, beyond which unsteady flow-induced vibrations are seen to occur.
Abstract-Short peptide ligands were synthesized to selectively functionalized microchips may not be ideal for use as capture spores using microcantilever arrays.
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