Quartz crystal microbalances for quantitative biosensing and characterizing protein multilayers.
The use of quartz crystal microbalances (QCMs) for quantitative biosensing and characterization of protein multilayers is demonstrated in three case studies. Monolayers of QCM-based affinity biosensors were investigated first. Layers of a thiol-containing synthetic peptide constituting an epitope of the foot-and-mouse-disease virus were formed on gold electrodes via self-assembly. The binding of specific antibodies to epitope-modified gold electrodes was detected for different concentrations of antibody solutions. Oligolayers were studied in a second set of experiments. Dextran hydrogels were modified by thrombin inhibitors. The QCM response was used in a competitive binding assay to identify inhibitors for thrombin at different concentrations. Multilayers of proteins formed by self-assembly of a biotin-conjugate and streptavidin were investigated next. The QCM frequency response was monitored as a function of layer thickness up to 20 protein layers. A linear frequency decay was observed with increasing thickness. The decay per layer remained constant, thus indicating perfect mass coupling to the substrate. Frequency changes a factor of four higher were obtained in buffer solution as compared to measurements in dry air. This indicates a significant incorporation of water (75% weight) in the protein layers. This water behaves like a solid concerning the shear mode coupling to the substrate. The outlook discusses briefly the need for controlled molecular engineering of overlayers for subsequent QCM analysis, and the importance of an additional multiparameter analysis with other transducer principles and with additional techniques of interface analysis to characterize the mechanical coupling of overlayers as biosensor coatings. A promising trend concerns the use of QCM-arrays for screening experiments.
Rickert
J
J
Institute of Physical and Theoretical Chemistry, University of Tübingen, Germany.
Brecht
A
A
Göpel
W
W
D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry; D002138:Calibration; D003460:Crystallization; D003911:Dextrans / Q000737:chemistry; D005782:Gels; D007700:Kinetics; D008832:Microchemistry; D010455:Peptides / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D011791:Quartz; D012709:Serum Albumin / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D014159:Transducers
D003911:Dextrans; D005782:Gels; D010455:Peptides; D011506:Proteins; D012709:Serum Albumin; D011791:Quartz; D012710:Serum Albumin, Bovine; D001710:Biotin