Development of biosensor-based assays to identify anti-infective oligosaccharides.
It is now well accepted that milk oligosaccharides can have a direct inhibitory effect on pathogenic microorganisms by interfering with their adhesion to human cells. Many free oligosaccharides from milk are considered to be soluble receptor analogs of epithelial cell surface carbohydrates and, thus, function as receptor decoys to which pathogens can bind instead of the host. In reality, there are few rapid methods to screen for such oligosaccharides, and much of the research in this area has centered on using human cell line models of infection that are time-consuming. Therefore, a quick and sensitive method is required for detecting the binding of microorganisms to milk oligosaccharides. Our study describes a number of biosensor-based methods to achieve these aims. Our approach involved the exposure of whole bacterial cells to the well-characterized human milk oligosaccharide, 2'-fucosyllactose, immobilized to a pretreated gold chip surface. The technique was validated by screening a range of pathogenic bacteria, including Campylobacter jejuni, to which 2'-fucosyllactose is known to bind. Where binding was detected, its specificity was confirmed by preincubation studies using unlabeled 2'-fucosyllactose. The techniques described represent a quick, cost-effective, and highly reproducible detection method for identifying anti-infective oligosaccharides.
Teagasc Food Research Centre, Moorepark, Fermoy, County Cork, Ireland.
Jonathan A
JA
Lane
Raj K
RK
Mehra
Stephen D
SD
Carrington
Rita M
RM
Hickey
D018829:Adhesins, Bacterial / Q000187:drug effects / Q000502:physiology; D000890:Anti-Infective Agents / Q000737:chemistry / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D016123:Campylobacter jejuni / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D008895:Milk, Human / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance / Q000379:methods; D014312:Trisaccharides / Q000737:chemistry* / Q000494:pharmacology*
D018829:Adhesins, Bacterial; D000890:Anti-Infective Agents; D009844:Oligosaccharides; D014312:Trisaccharides; C031420:2'-fucosyllactose