Lectin binding studies on a glycopolymer brush flow-through biosensor by localized surface plasmon resonance.
A localized surface plasmon resonance biosensor in a flow-through configuration was applied for investigating kinetics of lectin binding to surface-grafted glycopolymer brushes. Polycarbonate filter membranes with pore sizes of 400 nm were coated with a 114-nm thick gold layer and used as substrate for surface-initiated atom-transfer radical polymerization of a glycomonomer. These grafted from glycopolymer brushes were further modified with two subsequent enzymatic reactions on the surface to yield an immobilized trisaccharide presenting brush. Specific binding of lectins including Clostridium difficile toxin A receptor domain to the glycopolymer brush surface could be investigated in a microfluidic setup with flow-through of the analytes and transmission surface plasmon resonance spectroscopy. Graphical abstract Glycopolymer brushes serve as high affinity ligands for lectin and toxin interactions in a sensitive, disposable flow-through LSPR biosensor.
Laboratory for Biomaterials, Institute for Biotechnology and Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Pauwelsstr. 20, 52074, Aachen, Germany.|Fraunhofer Institute for Applied Polymer Research, Geiselbergstraße 69, 14476, Potsdam, Germany.
Ruben R
RR
Rosencrantz
Institute of Materials in Electrical Engineering 1, RWTH Aachen University, Sommerfeldstr. 24, 52074, Aachen, Germany.
Vu Hoa
VH
Nguyen
Leibniz Institute for Interactive Materials, Department for Macromolecular Materials and Surfaces at RWTH Aachen, Forckenbeckstr. 50, 52074, Aachen, Germany.
Hyunji
H
Park
Fraunhofer Institute for Applied Polymer Research, Geiselbergstraße 69, 14476, Potsdam, Germany.
Christine
C
Schulte
Leibniz Institute for Interactive Materials, Department for Macromolecular Materials and Surfaces at RWTH Aachen, Forckenbeckstr. 50, 52074, Aachen, Germany.|Chair of Polymer Materials and Polymer Technologies, University of Potsdam, Fraunhofer Institute for Applied Polymer Research, Geiselbergstraße 69, 14476, Potsdam, Germany.
Alexander
A
Böker
Institute of Materials in Electrical Engineering 1, RWTH Aachen University, Sommerfeldstr. 24, 52074, Aachen, Germany.
Uwe
U
Schnakenberg
Laboratory for Biomaterials, Institute for Biotechnology and Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Pauwelsstr. 20, 52074, Aachen, Germany. l.elling@biotec.rwth-aachen.de.
Lothar
L
Elling
D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D056656:Lab-On-A-Chip Devices; D037102:Lectins / Q000737:chemistry*; D011076:Polycarboxylate Cement / Q000737:chemistry*; D025941:Protein Interaction Mapping / Q000379:methods*; D011971:Receptors, Immunologic / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
C080487:Clostridium difficile enterotoxin A receptor; D037102:Lectins; D011076:Polycarboxylate Cement; D011971:Receptors, Immunologic; C005506:polycarbonate
Bacterial toxin; Biosensors; Glycopolymer brush; Glycosyltransferase; Localized surface plasmon resonance; Microfluidics