11 research outputs found
Development of a combined surface plasmon resonance/surface acoustic wave device for the characterization of biomolecules
It is known that acoustic sensor devices, if operated in liquid phase, are sensitive not just to the mass of the analyte but also to various other parameters, such as size, shape, charge and elastic constants of the analyte as well as bound and viscously entrained water. This can be used to extract valuable information about a biomolecule, particularly if the acoustic device is combined with another sensor element which is sensitive to the mass or amount of analyte only. The latter is true in good approximation for various optical sensor techniques. This work reports on the development of a combined surface plasmon resonance/surface acoustic wave sensor system which is designed for the investigation of biomolecules such as proteins or DNA. Results for the deposition of neutravidin and DNA are reported
Acoustic Array Biochip Combined with Allele-Specific PCR for Multiple Cancer Mutation Analysis in Tissue and Liquid Biopsy
[EN] Regular screening of point mutations is of importance to cancer management and treatment selection. Although techniques like next-generation sequencing and digital polymerase chain reaction (PCR) are available, these are lacking in speed, simplicity, and cost-effectiveness. The development of alternative methods that can detect the extremely low concentrations of the target mutation in a fast and cost-effective way presents an analytical and technological challenge. Here, an approach is presented where for the first time an allele-specific PCR (AS-PCR) is combined with a newly developed high fundamental frequency quartz crystal microbalance array as biosensor for the amplification and detection, respectively, of cancer point mutations. Increased sensitivity, compared to fluorescence detection of the AS-PCR amplicons, is achieved through energy dissipation measurement of acoustically ÂżlossyÂż liposomes binding to surface-anchored dsDNA targets. The method, applied to the screening of BRAF V600E and KRAS G12D mutations in spiked-in samples, was shown to be able to detect 1 mutant copy of genomic DNA in an excess of 104 wild-type molecules, that is, with a mutant allele frequency (MAF) of 0.01%. Moreover, validation of tissue and plasma samples obtained from melanoma, colorectal, and lung cancer patients showed excellent agreement with Sanger sequencing and ddPCR; remarkably, the efficiency of this AS-PCR/acoustic methodology to detect mutations in real samples was demonstrated to be below 1% MAF. The combined high sensitivity and technology-readiness level of the methodology, together with the ability for multiple sample analysis (24 array biochip), cost-effectiveness, and compatibility with routine workflow, make this approach a promising tool for implementation in clinical oncology labs for tissue and liquid biopsy.This work was supported by the European Union's Horizon H2020-FETOPEN-1-2016-2017 under grant agreement no. 737212 (CATCH-U-DNA).Naoumi, N.; Michaelidou, K.; Papadakis, G.; Simaiaki, AE.; FernĂĄndez DĂaz, R.; Calero-Alcarria, MDS.; Arnau Vives, A.... (2022). Acoustic Array Biochip Combined with Allele-Specific PCR for Multiple Cancer Mutation Analysis in Tissue and Liquid Biopsy. ACS Sensors. 7(2):495-503. https://doi.org/10.1021/acssensors.1c02245S4955037
On the Hydrodynamic Nature of DNA Acoustic Sensing
In this work we provide strong experimental
evidence for the hydrodynamic
nature of the acoustic wave/biomolecule interaction at a solid/liquid
interface. By using a wide range of DNAs of various sizes and by assuming
DNA attachment as discrete particles through a neutravidin/biotin
link, we prove experimentally that the acoustic ratio (dissipation/frequency)
is directly related to the moleculesâ intrinsic viscosity [η].
The relationship of [η] to the size and shape of biomolecules
is described in general and more specifically for linear dsDNA; equations
are derived linking the measured acoustic ratio to the number of dsDNA
base pairs for two acoustic sensors, the QCM and Love-wave devices
operating at a frequency of 35 and 155 MHz, respectively. Single-stranded
DNAs were also tested and shown to fit well to the equation derived
for the double-stranded molecules while new insight is provided on
their conformation on a surface. Other types of DNA are also shown
to fit the proposed model. The current work establishes a new way
of viewing acoustic sensor data and lays down the groundwork for a
surface technique where quantitative information can be obtained at
the nanometer scale regarding the shape and size, i.e., conformation
of biomolecules at an interface
Screening between normal and cancer human thyroid cells through comparative adhesion studies using the Quartz Crystal Microbalance
In this work, the Quartz Crystal Microbalance with Dissipation monitoring (QCM-D) was used to distinguish the dynamic cell adhesion behavior of human normal (Nthy) thyroid epithelial cells from poorly differentiated anaplastic carcinoma cells (ARO). The surfaces used to facilitate cell adhesion were bare titanium (Ti), gold (Au) and fibrinogen-coated gold (Fg-Au). The pattern of cell adhesion for both cell lines was that the largest acoustic signals were observed on Ti, followed by Au and last by Fg-Au; in addition, ARO cells always produced smaller acoustic signals than Nthy cells on the same surface and for the same number of cells in suspension. Moreover, the calculated acoustic ratio of energy dissipation over frequency change suggests a higher ability of Nthy cells to spread and potentially form more attachment points on the surface than the ARO cells, something observed in SEM images. Finally, we demonstrated that the application of two surfaces for cell adhesion experiments, one of which is Au and the other either Ti or Fg-Au, can discriminate with accuracy between the two particular cell types and potentially form a platform for differentiation between normal and cancer thyroid cell types. Keywords: QCM-D, Microscopy, Thyroid cancer cells, Gold, Titanium, Fibrinogen, Diagnostic
Hybrid Sensor Device for Simultaneous Surface Plasmon Resonance and Surface Acoustic Wave Measurements
International audienceSurface plasmon resonance (SPR) and Love wave (LW) surface acoustic wave (SAW) sensors have been established as reliable biosensing technologies for label-free, real-time monitoring of biomolecular interactions. This work reports the development of a combined SPR/LW-SAW platform to facilitate simultaneous optical and acoustic measurements for the investigation of biomolecules binding on a single surface. The systemâs output provides recordings of two acoustic parameters, phase and amplitude of a Love wave, synchronized with SPR readings. We present the design and manufacturing of a novel experimental set-up employing, in addition to the SPR/LW-SAW device, a 3D-printed plastic holder combined with a PDMS microfluidic cell so that the platform can be used in a flow-through mode. The system was evaluated in a systematic study of the optical and acoustic responses for different surface perturbations, i.e., rigid mass loading (Au deposition), pure viscous loading (glycerol and sucrose solutions) and protein adsorption (BSA). Our results provide the theoretical and experimental basis for future application of the combined system to other biochemical and biophysical studie
Extracting the Shape and Size of Biomolecules Attached to a Surface as Suspended Discrete Nanoparticles
The
ability to derive information on the conformation of surface
attached biomolecules by using simple techniques such as biosensors
is currently considered of great importance in the fields of surface
science and nanotechnology. Here we present a nanoshape sensitive
biosensor where a simple mathematical expression is used to relate
acoustic measurements to the geometrical features of a surface-attached
biomolecule. The underlying scientific principle is that the acoustic
ratio (Î<i>D</i>/Î<i>F</i>) is a measure
of the hydrodynamic volume of the attached entity, mathematically
expressed by its intrinsic viscosity [η]. A methodology is presented
in order to produce surfaces with discretely bound biomolecules where
their native conformation is maintained. Using DNA anchors we attached
a spherical protein (streptavidin) and a rod-shaped DNA (47bp) to
a quartz crystal microbalance (QCM) device in a suspended way and
predicted correctly through acoustic measurements their conformation,
i.e., shape and length. The methodology can be widely applied to draw
conclusions on the conformation of any biomolecule or nanoentity upon
specific binding on the surface of an acoustic wave device