| Products/Services Used | Details | Operation |
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| Peptide Synthesis | Following this step, the etched samples were immersed in 1 mM of 16-mercaptohexadecanoic acid (MHDA) thiol for 20 h. After the thiolation step, the biochips were sonicated in deoxygenated ethanol for 1 min and rinsed with degassed ethanol to remove unbound molecules. Thereafter, the thiolated samples were incubated for 30 min in the 0.4 M EDC (1-Ethyl-3-(3-Dimethylaminopropyl)Carbodiimnide) – 0.1 M NHS (N-HydroxySuccinimide) solution (1:1) to activate −COOH terminals. Following this procedure, the samples were immerged for 1 h in 0.1 mg/mL of warnericin RK AMP synthesized by GenScript Corporation, Piscataway, USA. The concentration of a peptide solution was chosen to match that of the L. pneumophila antibody solution (0.1 mg/mL). However, it is understood that a more comprehensive investigation will be necessary to optimize the biosensor performance. The functionalized samples were incubated with L. pneumophila, JR32 E. coli ATCC 25922, Bacillus subtilis ATCC 60514 (B. subtilis) and Pseudomonas fluorescens ATCC 13525 (P. fluorescens) at 106 CFU/mL, each, for 2 h. | Get A Quote |
Detection of pathogenic Legionella pneumophila by culture-based methods is not efficient in predicting outbreaks of the Legionnaires’ disease. The main problem is the relatively slow time-to-result and the inability of some culture media to support the growth of viable bacteria. One strategy to alleviate these issues is developing biosensors functionalized with mammalian antibodies designed to capture bacteria. However, mammalian antibodies are known to suffer from batch-to-batch variations, as well as limited stability, which reduce the consistent utility of antibody-based biosensors. In an attempt to address this problem, we investigated antimicrobial peptides (AMPs) for capture of L. pneumophila with G... More
