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label free biomolecular interaction

Francesco Chiavaioli, Pablo Zubiate, Ignacio Del Villar, Carlos Ruiz Zamarreño, Ambra Giannetti, Sara Tombelli, Cosimo Trono, Francisco J Arregui, Ignacio R Matias, Francesco Baldini
The advent of optical fibre-based biosensors combined with that of nanotechnologies has provided an opportunity for developing in situ, portable, lightweight, versatile and high-performance optical sensing platforms. We report on the generation of lossy mode resonances by the deposition of nm-thick metal oxide films on optical fibres, which makes it possible to measure precisely and accurately the changes in optical properties of the fibre-surrounding medium with very high sensitivity compared to other technology platforms, such as long period gratings or surface plasmon reso-nances, the gold standard in label-free and real-time biomolecular interaction analysis...
May 4, 2018: ACS Sensors
Matthew M Makowski, Cathrin Gräwe, Benjamin M Foster, Nhuong V Nguyen, Till Bartke, Michiel Vermeulen
Interaction proteomics studies have provided fundamental insights into multimeric biomolecular assemblies and cell-scale molecular networks. Significant recent developments in mass spectrometry-based interaction proteomics have been fueled by rapid advances in label-free, isotopic, and isobaric quantitation workflows. Here, we report a quantitative protein-DNA and protein-nucleosome binding assay that uses affinity purifications from nuclear extracts coupled with isobaric chemical labeling and mass spectrometry to quantify apparent binding affinities proteome-wide...
April 25, 2018: Nature Communications
Yu-Wen Su, Wei Wang
Surface plasmon resonance (SPR) has become a well-recognized label-free technique for measuring the binding kinetics between biomolecules since the invention of the first SPR-based immunosensor in 1980s. The most popular and traditional format for SPR analysis is to monitor the real-time optical signals when a solution containing ligand molecules is flowing over a sensor substrate functionalized with purified receptor molecules. In recent years, rapid development of several kinds of SPR imaging techniques have allowed for mapping the dynamic distribution of local mass density within single living cells with high spatial and temporal resolutions and reliable sensitivity...
April 12, 2018: Analytical and Bioanalytical Chemistry
M Gianneli, E Polo, H Lopez, V Castagnola, T Aastrup, K A Dawson
Nanomedicine, nanotargeting and nanotherapeutics have in the last few years faced several difficulties in translating the promising results obtained in vitro to an in vivo scenario. The origin of this discrepancy might be found in the lack of a detailed and realistic characterization of the biological surface of nanoparticles. Despite the capability to engineer nanomaterials with a great variety and a precise control of the surface functionalization, the targeting capability is lost when the nanoparticles are embedded in complex biological media, due to the formation of a biological layer (biomolecular corona)...
March 28, 2018: Nanoscale
Wen-Jing Bao, Jian Li, Jin Li, Qian-Wen Zhang, Yang Liu, Cai-Feng Shi, Xing-Hua Xia
A versatile and sensitive platform for label-free bioanalysis has been proposed on the basis of attenuated total reflection-surface enhanced infrared absorption spectroscopy (ATR-SEIRAS) using Au/ZnSe as the enhancement substrate that allows a wide spectral range down to 700 cm-1 . Au nanoparticles are stably deposited on the surface of a ZnSe prism due to the formation of Au-Se bonds via electroless deposition, and the enhancement factor of the resultant Au/ZnSe substrate is about 2 times larger than that of the commonly used Au/Si substrate...
March 20, 2018: Analytical Chemistry
Yuzhang Liang, Hui Zhang, Wenqi Zhu, Amit Agrawal, Henri Lezec, Lixia Li, Wei Peng, Yi Zou, Yanqing Lu, Ting Xu
With the development of advanced nanofabrication technologies over the past decade, plasmonic nanostructures have attracted wide attention for their potential in label-free biosensing applications. However, the sensing performance of nanostructured plasmonic sensors is primarily limited by the broad-line-width features with low peak-to-dip signal ratio in the extinction spectra that result from strong radiative damping. Here, we propose and systematically investigate the in-plane and out-of-plane dipolar interactions in an array of plasmonic nanoring resonators that are from the spatial combination of classic nanohole and nanodisk structures...
December 22, 2017: ACS Sensors
Lixia Li, Yuzhang Liang, Jianye Guang, Wenli Cui, Xinpu Zhang, Jean-Francois Masson, Wei Peng
We present a dual-resonance fiber surface plasmon resonance (SPR) sensor for biological analysis. The sensing element was fabricated by sequentially sputtering layers of indium tin oxide (ITO) (100 nm thickness) and Au (35 nm thickness) on the surface of an optical fiber. The refractive index dispersion effect of ITO material led to resonances in the near infrared and visible wavelength regions. The refractive index of ITO is larger than the optical fiber in visible spectral area (400 to 733nm), such that the structure is a typical Kretschmann configuration surface plasmon resonance sensor...
October 30, 2017: Optics Express
Samuel S Hinman, Kristy S McKeating, Quan Cheng
No abstract text is available yet for this article.
January 2, 2018: Analytical Chemistry
Saumya Prasad, Imon Mandal, Shubham Singh, Ashim Paul, Bhubaneswar Mandal, Ravindra Venkatramani, Rajaram Swaminathan
Electronic absorption spectra of proteins are primarily characterized over the ultraviolet region (185-320 nm) of the electromagnetic spectrum. While recent studies on peptide aggregates have revealed absorption beyond 350 nm, monomeric proteins lacking aromatic amino acids, disulphide bonds, and active site prosthetic groups are expected to remain optically silent beyond 250 nm. Here, in a joint theoretical and experimental investigation, we report the distinctive UV-Vis absorption spectrum between 250 nm [ε = 7338 M(-1) cm(-1)] and 800 nm [ε = 501 M(-1) cm(-1)] in a synthetic 67 residue protein (α3C), in monomeric form, devoid of aromatic amino acids...
August 1, 2017: Chemical Science
Volker Gatterdam, Andreas Frutiger, Klaus-Peter Stengele, Dieter Heindl, Thomas Lübbers, Janos Vörös, Christof Fattinger
Focal molography is a next-generation biosensor that visualizes specific biomolecular interactions in real time. It transduces affinity modulation on the sensor surface into refractive index modulation caused by target molecules that are bound to a precisely assembled nanopattern of molecular recognition sites, termed the 'mologram'. The mologram is designed so that laser light is scattered at specifically bound molecules, generating a strong signal in the focus of the mologram via constructive interference, while scattering at nonspecifically bound molecules does not contribute to the effect...
November 2017: Nature Nanotechnology
Ugur Aygun, Oguzhan Avci, Elif Seymour, Hakan Urey, M Selim Ünlü, Ayca Yalcin Ozkumur
Fluorescence based microarray detection systems provide sensitive measurements; however, variation of probe immobilization and poor repeatability negatively affect the final readout, and thus quantification capability of these systems. Here, we demonstrate a label-free and high-throughput optical biosensor that can be utilized for calibration of fluorescence microarrays. The sensor employs a commercial flatbed scanner, and we demonstrate transformation of this low cost (∼100 USD) system into an Interferometric Reflectance Imaging Sensor through hardware and software modifications...
October 27, 2017: ACS Sensors
Erhan Süleymanoğlu
Despite the significant efforts towards applying improved non-destructive and label-free measurements of biomolecular structures of lipid-based gene delivery vectors, little is achieved in terms of their structural relevance in gene transfections. Better understanding of structure-activity relationships of lipid-DNA complexes and their gene expression efficiencies thus becomes an essential issue. Raman scattering offers a complimentary measurement technique for following the structural transitions of both DNA and lipid vesicles employed for their transfer...
December 2017: Journal of Biological Inorganic Chemistry: JBIC
José Juan-Colás, Steven Johnson, Thomas F Krauss
The monitoring of biomolecular interactions is a key requirement for the study of complex biological processes and the diagnosis of disease. Technologies that are capable of providing label-free, real-time insight into these interactions are of great value for the scientific and clinical communities. Greater understanding of biomolecular interactions alongside increased detection accuracy can be achieved using technology that can provide parallel information about multiple parameters of a single biomolecular process...
September 7, 2017: Sensors
Achyut J Raghavendra, Kristofer Fritz, Sherleen Fu, Jared M Brown, Ramakrishna Podila, Jonathan H Shannahan
Ball-milling utilizes mechanical stress to modify properties of carbon nanotubes (CNTs) including size, capping, and functionalization. Ball-milling, however, may introduce structural defects resulting in altered CNT-biomolecule interactions. Nanomaterial-biomolecule interactions result in the formation of the biocorona (BC), which alters nanomaterial properties, function, and biological responses. The formation of the BC is governed by the nanomaterial physicochemical properties and the physiological environment...
August 16, 2017: Scientific Reports
Massimo Rippa, Riccardo Castagna, Marianna Pannico, Pellegrino Musto, Giorgia Borriello, Rubina Paradiso, Giorgio Galiero, Sergio Bolletti Censi, Jun Zhou, Joseph Zyss, Lucia Petti
The development of fast and ultrasensitive methods to detect bacterial pathogens at low concentrations is of high relevance for human and animal health care and diagnostics. In this context, surface-enhanced Raman scattering (SERS) offers the promise of a simplified, rapid, and high-sensitive detection of biomolecular interactions with several advantages over previous assay methodologies. In this work, we have conceived reproducible SERS nanosensors based on tailored multilayer octupolar nanostructures which can combine high enhancement factor and remarkable molecular selectivity...
July 28, 2017: ACS Sensors
Lorena Mendive-Tapia, Ramon Subiros-Funosas, Can Zhao, Fernando Albericio, Nick D Read, Rodolfo Lavilla, Marc Vendrell
Fluorescent peptides are valuable tools for live-cell imaging because of the high specificity of peptide sequences for their biomolecular targets. When preparing fluorescent versions of peptides, labels must be introduced at appropriate positions in the sequences to provide suitable reporters while avoiding any impairment of the molecular recognition properties of the peptides. This protocol describes the preparation of the tryptophan (Trp)-based fluorogenic amino acid Fmoc-Trp(C2-BODIPY)-OH and its incorporation into peptides for live-cell fluorescence imaging-an approach that is applicable to most peptide sequences...
August 2017: Nature Protocols
Danlin Yang, Ajit Singh, Helen Wu, Rachel Kroe-Barrett
Label-free optical biosensors are powerful tools in drug discovery for the characterization of biomolecular interactions. In this study, we describe the use of four routinely used biosensor platforms in our laboratory to evaluate the binding affinity and kinetics of ten high-affinity monoclonal antibodies (mAbs) against human proprotein convertase subtilisin kexin type 9 (PCSK9). While both Biacore T100 and ProteOn XPR36 are derived from the well-established Surface Plasmon Resonance (SPR) technology, the former has four flow cells connected by serial flow configuration, whereas the latter presents 36 reaction spots in parallel through an improvised 6 x 6 crisscross microfluidic channel configuration...
April 17, 2017: Journal of Visualized Experiments: JoVE
Jordan Boutilier, Hong M Moulton
Surface plasmon resonance (SPR) is a physical process that allows label-free and real-time detection of biomolecular interactions. SPR provides a rapid and quantitative method for studying interactions of macromolecules such as proteins and nucleic acids. Antisense Morpholino oligomers are widely used to regulate gene expression and the US FDA has approved a Morpholino drug for treatment of Duchenne muscular dystrophy. Here, we describe an antibody-free, label-free, high-throughput, and walk-away SPR method for quantification of Morpholino compounds extracted from biological specimens...
2017: Methods in Molecular Biology
Jakob Wallner, Gabriele Lhota, Markus Schosserer, Karola Vorauer-Uhl
Non-fluidic bio-layer interferometry (BLI) has rapidly become a standard tool for monitoring almost all biomolecular interactions in a label-free, real-time and high-throughput manner. High-efficiency screening methods which measure the kinetics of liposomes with a variety of compounds require the immobilization of liposomes. In this work, a method is described for immobilizing liposomes for interaction studies, based on the biophysical principles of this biosensor platform. The immobilization approach includes the loading of DSPE-PEG(2000) -biotin containing sterically stabilized micelles (SSMs) which are restructured in a buffer change step, resulting in an accessible substrate for liposome immobilization...
June 1, 2017: Colloids and Surfaces. B, Biointerfaces
Charalambos Kallepitis, Mads S Bergholt, Manuel M Mazo, Vincent Leonardo, Stacey C Skaalure, Stephanie A Maynard, Molly M Stevens
The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment...
March 22, 2017: Nature Communications
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