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We describe a highly sensitive, substrate-specific, label-free, and multiplexed protease assay which reports proteases activities as an optical bar chart, allowing test results to be easily assessed by laymen with the naked eye. First, an oligopeptide microarray having six rows of immobilized oligopeptides, with well-controlled orientations and concentration gradients, is immersed in a buffer solution containing proteases. Then, a thin layer liquid crystal is supported on the microarray to transduce the oligopeptide cleavage event into an optical bar chart of different colors and lengths. This type of optical bar chart provides very rich information such as protease concentration, incubation time, surface densities of oligopeptides, etc. Both trypsin and chymotrypsin can be detected by using this assay within 3 h. The capability of the multiplexed protease assay opens up possibilities for detecting toxins such as botulinum neurotoxins which are known to cleave proteins and affect the docking and fusing synaptic vesicles.

In this article, we report a quartz crystal microbalance (QCM)-based detection method which allows the identification and quantification of two neonicotinoid pesticides, imidacloprid and thiacloprid, in aqueous solutions and celery juice. To achieve high selectivity, molecular imprinted monolayers (MIMs), which can either recognize 1 [micro]M of imidacloprid or 1 [micro]M of thiacloprid, are prepared from alkanethiols self-assembled on QCM sensor chips with preadsorbed templates (either imidacloprid or thiacloprid). Our experimental results show that the detection limit can be improved by using alkanethiols having longer hydrocarbon chains. For example, MIMs prepared from hexadecanethiol have dissociation constants 2-5 times smaller than those prepared from octanethiol. To detect two neonicotinoids in vegetable samples simultaneously, we also develop a new type of MIM with two different templates. A single QCM decorated with this MIM can respond to 10 [micro]M of imidacloprid and 10 [micro]M thiacloprid in celery juice in a real-time manner.

Author(s): Jianzhong He [1]; Kirsti M. Ritalahti [1]; Kun-Lin Yang [1]; Stephen S. Koenigsberg [2]; Frank E. Löffler (corresponding author) [1, 3] Tetrachloroethene (PCE) and trichloroethene (TCE) are ideal solvents [...]