Characterisation of an antibody coated microcantilever as a potential immuno-based biosensor

Grogan, Catherine; Raiteri, Roberto; O'Connor, Gerard M.; Glynn, Thomas J.; Cunningham, Vincent; Kane, M; Charlton, Martin D. B.; Leech, Dónal

doi:10.1016/s0956-5663(01)00276-7

Cited by 118 publications

(80 citation statements)

References 15 publications

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“…Similar behavior has been observed in previous studies measuring macro-molecular interactions such as with protein [3,4] and DNA [5,6]. Microcantilever responses have also proven useful in label-free biodetection [6], quantitative binding assays [7,8] and proteomic analysis [9].…”

Section: Introductionsupporting

confidence: 76%

Measurement of Temperature Induced Unfolding of DNA Hairpins by Microcantilever Sensors

Ng¹,

Dowell²,

Kar³

et al. 2013

OJAB

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The technical feasibility of monitoring DNA melting and cooling transitions using a microcantilever substrate has been demonstrated and these results were compared with those measured in solution by circular dichroism. DNA hairpins have been immobilized on the surface of gold-coated microcantilever surfaces and their DNA melting and cooling transitions were monitored by nanomechanical deflections. The hairpins comprised of a 16 base-pair GACA repeat motif stem duplex with a 29 nucleotide variable region. Microcantilever deflection profiles, measured by the microcantilever response as a function of temperature, were unique to different hairpins indicative of the molecules' general stability and denaturation characteristics. The major melting and cooling transition temperatures for all three immobilized oligonucleotides were between 41˚C -52˚C. The composition and flexibility of the DNA stem loops were shown to influence the thermal transitions.

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Section: Introductionsupporting

confidence: 76%

Measurement of Temperature Induced Unfolding of DNA Hairpins by Microcantilever Sensors

Ng¹,

Dowell²,

Kar³

et al. 2013

OJAB

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“…Further advances in nanomechanics allowed the use of microcantilevers for the detection of biomarkers and the transduction of DNA hybridisation into a nanomechanical response [9][10][11], along with the use of silicon cantilevers for the detection of volatile organic compounds [5]. Silicon cantilever technology also uses microcantilevers for the detection of malignant melanoma [12,13].…”

Section: Introductionmentioning

confidence: 99%

Microcantilever arrays functionalised with spiropyran photoactive moieties as systems to measure photo-induced surface stress changes

Grogan¹,

Florea

Koprivica

et al. 2016

Sensors and Actuators B: Chemical

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Herein we investigate the feasibility of detecting photo-induced surface stress changes using the deflection response of cantilevers. For this purpose, silicon microcantilevers have been functionalised with spiropyran photochromic molecules, using both a monolayer and a polymeric brushes approach. Upon ultraviolet light irradiation, the spiropyran unit is converted to the merocyanine form due to the photo-induced cleavage of the C spiro -O bond. The two forms of the molecule have dramatically different charge, polarity and molecular conformations. This makes spiropyrans an ideal system to study the correlation between photo-induced molecular changes and corresponding changes in surface stress. Our investigations include monitoring the changes in static cantilever deflection, and consequently, surface stress of the spiropyran functionalised cantilevers on exposure to ultraviolet light. Cantilever deflection data reveals that ultraviolet induced conformational changes in the spiropyran moiety cause a change in compressive surface 2 stress and this varies with the type of functionalisation method implemented. The change in surface stress response from the spiropyran polymer brushes functionalised cantilevers gives an average surface stress change of 98 Nm -1 (n = 8) while the spiropyran monolayer coated cantilevers have an average surface stress change of about 446 Nm -1 (n = 24) upon irradiation with UV light.

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“…Single microcantilevers are generally known to be susceptible to spurious deflections because of temperature changes, chemical interaction of a coated cantilever with the liquid environment, and slow rearrangement of molecules in the protein multilayer (10,12,26,27). As a result, a base-line drift can be observed during static-mode measurements.…”

Section: Resultsmentioning

confidence: 99%

A label-free immunosensor array using single-chain antibody fragments

Backmann

Zahnd

Huber

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

264

183

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We report a microcantilever-based immunosensor operated in static deflection mode with a performance comparable with surface plasmon resonance, using single-chain Fv (scFv) antibody fragments as receptor molecules. As a model system scFv fragments with specificity to two different antigens were applied. We introduced a cysteine residue at the C terminus of each scFv construct to allow covalent attachment to gold-coated sensor interfaces in directed orientation. Application of an array enabled simultaneous deflection measurements of sensing and reference cantilevers. The differential deflection signal revealed specific antigen binding and was proportional to the antigen concentration in solution. Using small, oriented scFv fragments as receptor molecules we increased the sensitivity of microcantilevers to Ϸ1 nM.cantilever arrays ͉ nanomechanics ͉ proteomics M icrocantilever-based sensors have attracted much interest as devices for fast and reliable detection of small amounts of molecules in air and solution. Over the last few years the application of the cantilever sensor concept was extended to the measurements of biocompounds in solution, resulting in a versatile biosensor (1, 2). Because of its label-free detection principle and small size, this kind of biosensor is advantageous for diagnostic applications, disease monitoring, and research in genomics or proteomics (3, 4). Multicantilever arrays would enable the detection of several analytes simultaneously.The main principle of the cantilever static mode is the transduction of the molecular interaction between analyte and receptors, immobilized as a layer on one surface of a cantilever, into a nanomechanical motion of the cantilever. Biomolecular interactions taking place on a solid-state interface produce a change in surface stress, because of changes in molecular configuration and intermolecular crowding (5). This process results in bending of the cantilever. Microcantilever-based biosensors operated in static mode have been successfully applied for the detection of various molecular interactions such as ssDNA-ssDNA (5-7) or protein-DNA (8, 9). Interactions between proteins were detected with cantilever-based immunosensors, where an antigen was recognized by its cognate antibody randomly immobilized on the sensor surface (10-12).The most critical step in preparation of any immunosensor is the immobilization of capture molecules on the support, a process where the orientation of the antigen-binding sites toward the analyte in solution plays a key role. Immunoglobulins can be either adsorbed on gold directly (10, 12) or attached covalently to the surface modified with hetero-bifunctional self-assembled monolayers of alkylthiols (11). However, these approaches produce a layer of randomly oriented antibody molecules on the cantilever surface, thereby generating conformational heterogeneity and inactive receptor molecules (13,14).As previously shown (13,(15)(16)(17)(18), the sensitivity of immunosensors can be improved by both maximizing the degree of functional ...

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Characterisation of an antibody coated microcantilever as a potential immuno-based biosensor

Cited by 118 publications

References 15 publications

Measurement of Temperature Induced Unfolding of DNA Hairpins by Microcantilever Sensors

Measurement of Temperature Induced Unfolding of DNA Hairpins by Microcantilever Sensors

Microcantilever arrays functionalised with spiropyran photoactive moieties as systems to measure photo-induced surface stress changes

A label-free immunosensor array using single-chain antibody fragments

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