Bacteria repellent surfaces and antibody-based coatings for bacterial assays have shown a growing demand in the field of biosensors, and have crucial importance in the design of biomedical devices. However, in-depth investigations and comparisons of possible solutions are still missing. The optical waveguide lightmode spectroscopy (OWLS) technique offers label-free, non-invasive, in situ characterization of protein and bacterial adsorption. Moreover, it has excellent flexibility for testing various surface coatings. Here, we describe an OWLS-based method supporting the development of bacteria repellent surfaces and characterize the layer structures and affinities of different antibody-based coatings for bacterial assays. In order to test nonspecific binding blocking agents against bacteria, OWLS chips were coated with bovine serum albumin (BSA), I-block, PAcrAM-g-(PMOXA, NH2, Si), (PAcrAM-P) and PLL-g-PEG (PP) (with different coating temperatures), and subsequent Escherichia coli adhesion was monitored. We found that the best performing blocking agents could inhibit bacterial adhesion from samples with bacteria concentrations of up to 107 cells/mL. Various immobilization methods were applied to graft a wide range of selected antibodies onto the biosensor’s surface. Simple physisorption, Mix&Go (AnteoBind) (MG) films, covalently immobilized protein A and avidin–biotin based surface chemistries were all fabricated and tested. The surface adsorbed mass densities of deposited antibodies were determined, and the biosensor;s kinetic data were evaluated to divine the possible orientations of the bacteria-capturing antibodies and determine the rate constants and footprints of the binding events. The development of affinity layers was supported by enzyme-linked immunosorbent assay (ELISA) measurements in order to test the bacteria binding capabilities of the antibodies. The best performance in the biosensor measurements was achieved by employing a polyclonal antibody in combination with protein A-based immobilization and PAcrAM-P blocking of nonspecific binding. Using this setting, a surface sensitivity of 70 cells/mm2 was demonstrated.
A modern gyógyászat egyre inkább igényli a páciensközeli diagnosztikát a terápiás döntések felgyorsítása vagy akár a laboratóriumi vizsgálatok kiváltása érdekében. Ehhez olyan megbízható, költséghatékony bioanalitikai mérőrendsze-rek szükségesek, amelyek mintegy mikrolaboratóriumként, integráltan tartalmazzák a biomolekuláris felismerő, érzé-kelő és jelfeldolgozó, illetve mikrofl uidikai mintapreparációs modulokat. Ezek a jórészt mikro-és nanotechnológiai fabrikációs módszerekkel kialakított Lab-on-a-chip rendszerek új távlatokat nyithatnak a gyógyászati ellátás láncola-tában, hiszen alkalmasak lehetnek speciális betegségmarkerek vagy azok kombinációinak egy csepp mintából történő, kvantitatív, nagy pontosságú és azonnali kimutatására is. Ennek megfelelően a mérőműszerekkel és az alkalmazott analitikai módszerekkel szembeni legfőbb követelmények a magas szelektivitás, alacsony alsó mérési határ, rövid vá-laszidő, integrálhatóság egészségügyi informatikai rendszerekbe. Mindez lehetőséget teremthet a vizsgálatokhoz szükséges hierarchikus lánc rövidítésére is, ezáltal gyökeresen átalakíthatja a laboratóriumi diagnosztikát, új helyzetet teremtve a terápiás intervencióban. Orv. Hetil., 2015, 156(52), 2096-2102.Kulcsszavak: betegágy melletti diagnosztika, in vitro diagnosztika, laboratórium egy chipen, mikrofl uidika, bioszenzor, biomarker, információs rendszer Lab-on-a-chip systems in the point-of-care diagnosticsThe need in modern medicine for near-patient diagnostics being able to accelerate therapeutic decisions and possibly replacing laboratory measurements is signifi cantly growing. Reliable and cost-effective bioanalytical measurement systems are required which -acting as a micro-laboratory -contain integrated biomolecular recognition, sensing, signal processing and complex microfl uidic sample preparation modules. These micro-and nanofabricated Lab-ona-chip systems open new perspectives in the diagnostic supply chain, since they are able even for quantitative, highprecision and immediate analysis of special disease specifi c molecular markers or their combinations from a single drop of sample. Accordingly, crucial requirements regarding the instruments and the analytical methods are the high selectivity, extremely low detection limit, short response time and integrability into the healthcare information networks. All these features can make the hierarchical examination chain shorten, and revolutionize laboratory diagnostics, evolving a brand new situation in therapeutic intervention.Keywords: point-of-care diagnostics, in vitro diagnostics, Lab-on-a-chip, microfl uidics, biosensor, biomarker, information system Szabó, B., Borbíró, A., Fürjes, P. [Lab-on-a-chip systems in the point-of-care diagnostics]. Orv. Hetil., 2015, 156(52), 2096-2102. (Beérkezett: 2015 elfogadva: 2015. október Rövidítések COC = cikloolefi n kopolimer; HIS = (hospital information system) kórházi információs rendszer; IVD = in vitro diagnostics; LIS = (laboratory information system) laboratóriumi információs rendszer; LOC = (Lab-on-a...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.