Bacterial capture by peptide-mimetic oligoacyllysine surfaces.
Most procedures for detecting pathogens in liquid media require an initial concentration step. However, poor recovery efficiencies of conventional methods, such as filtration, often lead to low sensitivity. Here, we describe a strategy for concentrating bacteria using their binding affinity for an oligoacyllysine (OAK), a novel peptide-mimetic antimicrobial compound. We show that the resin-linked OAK (ROAK) efficiently captures a variety of pathogens in different media, upon brief incubation with ROAK beads or after continuous flow through a ROAK-packed column. Using Escherichia coli expressing green fluorescent protein, we show that binding occurs rapidly during incubation and persists after filtration as visualized by confocal microscopy. The high binding affinity of bacteria was confirmed by surface plasmon resonance technology using an OAK-linked chip. ROAK-bound bacteria remained viable and were readily identifiable by real-time PCR after ethanol elution. A single ROAK bead is estimated to capture about 3,000 bacterial cells in culture medium, in contaminated saline or tap water. ROAK beads can be regenerated for multiple uses after brief ethanol treatment. Collectively, the data support the notion that OAK-based coating of polymeric surfaces might represent a useful means for medium filtration as well as for concentration of bacteria.
Rotem
Shahar
S
Department of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa, Israel.
Raz
Nili
N
Kashi
Yechezkel
Y
Mor
Amram
A
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects / Q000378:metabolism; D018407:Bacterial Physiological Phenomena; D001431:Bacteriological Techniques / Q000295:instrumentation* / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D020349:Surface Plasmon Resonance
D000900:Anti-Bacterial Agents; D049452:Green Fluorescent Proteins