共查询到20条相似文献,搜索用时 62 毫秒
1.
Lee R Kim J Kim SY Jang SM Lee SM Choi IH Park SW Shin JS Yoo KH 《Lab on a chip》2012,12(13):2377-2384
Label-free cell-based assays have emerged as a promising means for high-throughput screening. Most label-free sensors are based on impedance measurements that reflect the passive electrical properties of cells. Here we introduce a capacitance-based assay that measures the dielectric constant (capacitance) of biological cells, and demonstrate the feasibility of analyzing endocytosis and screening chemotherapeutic agents with this assay. Endocytosis induces a change in the zeta potential, leading to a change in the dielectric constant which enables real-time endocytosis monitoring using the capacitance sensor. Additionally, since the dielectric constant is proportional to cell radius and cell volume, cell viability can be estimated from the change in capacitance. Therefore, the capacitance sensor array can also be used for cytotoxicity testing for large-scale chemotherapeutic screening. 相似文献
2.
Surface-immobilized protein species deposited in the microarray format have been detected by time-of-flight secondary ion mass spectrometry and by scanning Kelvin nanoprobe. The former method was used to examine the nature of protein deposition on amine-coated glass slides and gold substrates in preparation for Kelvin measurements. Both gallium and SF(5)(+) ion sources were employed to produce positive and negative ion spectra of amino acids and polypeptides. Scanning Kelvin technology has been used to detect antibody-antigen interactions in a label-free protocol through measurement of the surface potential of the biochemical pair on indium tin oxide, amine-treated slides and gold substrates. The results show that good inter-spot reproducibility can be achieved and that deposited areas can be examined for homogeneity at 100 nm resolution. This work represents the first report on surface potential detection in protein microarray technology. 相似文献
3.
Carbohydrate recognition is clearly present throughout nature, playing a major role in the initial attachment of one biological
entity to another. The important question is whether these prevalent interactions could provide a real suitable alternative
to the use of antibodies or nucleic acid for detection and identification. Currently, examples of carbohydrates being employed
in biological detection systems are limited. The challenges of using carbohydrate recognition for detection mainly come from
the weak affinity of carbohydrate–protein interactions, the lack of versatile carbohydrate scaffolds with well-defined structures,
and the less developed high-information-content, real-time, and label-free assay technology. In this review, we focus on discussing
the characteristics of carbohydrate–protein interactions in nature and the methods for carbohydrate immobilization based on
surface coupling chemistry in terms of their general applicability for developing carbohydrate- and lectin-based label-free
sensors. Furthermore, examples of innovative design of multivalent carbohydrate–protein interactions for sensor applications
are given. We limit our review to show the feasibility of carbohydrate and lectin as recognition elements for label-free sensor
development in several representative cases to formulate a flexible platform for their use as recognition elements for real-world
biosensor applications. 相似文献
4.
Present and future of surface plasmon resonance biosensors 总被引:22,自引:0,他引:22
Homola J 《Analytical and bioanalytical chemistry》2003,377(3):528-539
Surface plasmon resonance (SPR) biosensors are optical sensors exploiting special electromagnetic waves—surface plasmon-polaritons—to probe interactions between an analyte in solution and a biomolecular recognition element immobilized on the SPR sensor surface. Major application areas include detection of biological analytes and analysis of biomolecular interactions where SPR biosensors provide benefits of label-free real-time analytical technology. This paper reviews fundamentals of SPR affinity biosensors and discusses recent advances in development and applications of SPR biosensors. 相似文献
5.
A label-free protein microfluidic array for immunoassays based on the combination of imaging ellipsometry and an integrated microfluidic system is presented. Proteins can be patterned homogeneously on substrate in array format by the microfluidic system simultaneously. After preparation, the protein array can be packed in the microfluidic system which is full of buffer so that proteins are not exposed to denaturing conditions. With simple microfluidic channel junction, the protein microfluidic array can be used in serial or parallel format to analyze single or multiple samples simultaneously. Imaging ellipsometry is used for the protein array reading with a label-free format. The biological and medical applications of the label-free protein microfluidic array are demonstrated by screening for antibody-antigen interactions, measuring the concentration of the protein solution and detecting five markers of hepatitis B. 相似文献
6.
The appeal of microarray technology is the possibility of large-scale parallel determination of a variety of variables simultaneously. Hence, microarray technologies attract the interest of both the scientific and business worlds alike. High-throughput screening has been the major focus of the utilization of microarray technologies in recent years, and has provided the strong driving force for developments in this field. DNA chip and biochip technologies have been developed as a consequence of worldwide activity in genome research. This review focuses on microarray-based analysis and emphasizes some of its principal constraints, especially detection limits. 相似文献
7.
Microarray formats have become an important tool for parallel (or multiplexed) monitoring of biomolecular interactions. Surface-immobilized probes like oligonucleotides, cDNA, proteins, or antibodies can be used for the screening of their complementary targets, covering different applications like gene or protein expression profiling, analysis of point mutations, or immunodiagnostics. Numerous reviews have appeared on this topic in recent years, documenting the intriguing progress of these miniaturized assay formats. Most of them highlight all aspects of microarray preparation, surface chemistry, and patterning, and try to give a systematic survey of the different kinds of applications of this new technique. This review places the emphasis on optical technologies for microarray analysis. As the fluorescent read out of microarrays is dominating the field, this topic will be the focus of the review. Basic principles of labeling and signal amplification techniques will be introduced. Recent developments in total internal reflection fluorescence, resonance energy transfer assays, and time-resolved imaging are addressed, as well as non-fluorescent imaging methods. Finally, some label-free detection modes are discussed, such as surface plasmon microscopy or ellipsometry, since these are particularly interesting for microarray development and quality control purposes. 相似文献
8.
Increasing the sensitivity in DNA microarray hybridization can significantly enhance the capability of microarray technology
for a wide range of research and clinical diagnostic applications, especially for those with limited sample biomass. To address
this issue, using reverse microemulsion method and surface chemistry, a novel class of homogenous, photostable, highly fluorescent
streptavidin-functionalized silica nanoparticles was developed, in which Alexa Fluor 647 (AF647) molecules were covalently
embedded. The coating of bovine serum albumin on the resultant fluorescent particles can greatly eliminate nonspecific background
signal interference. The thus-synthesized fluorescent nanoparticles can specifically recognize biotin-labeled target DNA hybridized
to the microarray via streptavidin–biotin interaction. The response of this DNA microarray technology exhibited a linear range
within 0.2 to 10 pM complementary DNA and limit of detection of 0.1 pM, enhancing microarray hybridization sensitivity over
tenfold. This promising technology may be potentially applied to other binding events such as specific interactions between
proteins. 相似文献
9.
Proll G Steinle L Pröll F Kumpf M Moehrle B Mehlmann M Gauglitz G 《Journal of chromatography. A》2007,1161(1-2):2-8
The classical approach of high-content screening (HCS) is based on multiplexed, functional cell-based screening and combines several analytical technologies that have been used before separately to achieve a better level of automation (scale-up) and higher throughput. New HCS methods will help to overcome the bottlenecks, e.g. in the present development chain for lead structures for the pharmaceutical industry or during the identification and validation process of new biomarkers. In addition, there is a strong need in analytical and bioanalytical chemistry for functional high-content assays which can be provided by different hyphenated techniques. This review discusses the potential of a label-free optical biosensor based on reflectometric interference spectroscopy (RIfS) as a bridging technology for different HCS approaches. Technical requirements of RIfS are critically assessed by means of selected applications and compared to the performance characteristics of surface plasmon resonance (SPR) which is currently the leading technology in the area of label-free optical biosensors. 相似文献
10.
Bosco FG Hwu ET Chen CH Keller S Bache M Jakobsen MH Hwang IS Boisen A 《Lab on a chip》2011,11(14):2411-2416
Sensors are crucial in many daily operations including security, environmental control, human diagnostics and patient monitoring. Screening and online monitoring require reliable and high-throughput sensing. We report on the demonstration of a high-throughput label-free sensor platform utilizing cantilever based sensors. These sensors have often been acclaimed to facilitate highly parallelized operation. Unfortunately, so far no concept has been presented which offers large datasets as well as easy liquid sample handling. We use optics and mechanics from a DVD player to handle liquid samples and to read-out cantilever deflection and resonant frequency. Also, surface roughness is measured. When combined with cantilever deflection the roughness is discovered to hold valuable additional information on specific and unspecific binding events. In a few minutes, 30 liquid samples can be analyzed in parallel, each by 24 cantilever-based sensors. The approach was used to detect the binding of streptavidin and antibodies. 相似文献
11.
A convenient strategy for the preparation of label-free selective protein biomarker sensors is presented. After formation of a ferrocene terminated monolayer on a gold electrode using thiol chemistry, grafting of an antifouling phenylalanine layer onto the preformed ferrocene monolayer is achieved by diazonium chemistry. This provides a platform for receptor (anti-BSA or anti-CRP) immobilization as well an antifouling interface. Specific target binding leads to attenuation of the voltammetric faradaic signal of the underlying ferrocene group. Thereby, label-free, selective, voltammetric detection is achieved with highly competitive limits of detection in the low (≤4) picomolar range for both BSA and CRP. 相似文献
12.
An investigation of the mechanisms of electronic sensing of protein adsorption on carbon nanotube devices 总被引:5,自引:0,他引:5
Chen RJ Choi HC Bangsaruntip S Yenilmez E Tang X Wang Q Chang YL Dai H 《Journal of the American Chemical Society》2004,126(5):1563-1568
It has been reported that protein adsorption on single-walled carbon nanotube field effect transistors (FETs) leads to appreciable changes in the electrical conductance of the devices, a phenomenon that can be exploited for label-free detection of biomolecules with a high potential for miniaturization. This work presents an elucidation of the electronic biosensing mechanisms with a newly developed microarray of nanotube "micromat" sensors. Chemical functionalization schemes are devised to block selected components of the devices from protein adsorption, self-assembled monolayers (SAMs) of methoxy(poly(ethylene glycol))thiol (mPEG-SH) on the metal electrodes (Au, Pd) and PEG-containing surfactants on the nanotubes. Extensive characterization reveals that electronic effects occurring at the metal-nanotube contacts due to protein adsorption constitute a more significant contribution to the electronic biosensing signal than adsorption solely along the exposed lengths of the nanotubes. 相似文献
13.
14.
Piezoelectric microelectromechanical resonant sensors for chemical and biological detection 总被引:1,自引:0,他引:1
Piezoelectric microelectromechanical systems (MEMS) resonant sensors, known for their excellent mass resolution, have been studied for many applications, including DNA hybridization, protein-ligand interactions, and immunosensor development. They have also been explored for detecting antigens, organic gas, toxic ions, and explosives. Most piezoelectric MEMS resonant sensors are acoustic sensors (with specific coating layers) that enable selective and label-free detection of biological events in real time. These label-free technologies have recently garnered significant attention for their sensitive and quantitative multi-parameter analysis of biological systems. Since piezoelectric MEMS resonant sensors do more than transform analyte mass or thickness into an electrical signal (e.g., frequency and impedance), special attention must be paid to their potential beyond microweighing, such as measuring elastic and viscous properties, and several types of sensors currently under development operate at different resonant modes (i.e., thickness extensional mode, thickness shear mode, lateral extensional mode, flexural mode, etc.). In this review, we provide an overview of recent developments in micromachined resonant sensors and activities relating to biochemical interfaces for acoustic sensors. 相似文献
15.
Changqing Yi Cheuk-Wing Li Huayang Fu Mingliang Zhang Suijian Qi Ning-Bew Wong Shuit-Tong Lee Mengsu Yang 《Analytical and bioanalytical chemistry》2010,397(7):3143-3150
Patterning is of paramount importance in many areas of modern science and technology. As a good candidate for novel nanoscale optoelectronics and miniaturized molecule sensors, vertically aligned silicon nanowire (SiNW) with controllable location and orientation is highly desirable. In this study, we developed an effective procedure for the fabrication of vertically aligned SiNW arrays with micro-sized features by using single-step photolithography and silver nanoparticle-induced chemical etching at room temperature. We demonstrated that the vertically aligned SiNW arrays can be used as a platform for label-free DNA detection using surface-enhanced Raman spectroscopy (SERS), where the inherent “fingerprint” SERS spectra allows for the differentiation of closely related biospecies. Since the SiNW array patterns could be modified by simply varying the mask used in the photolithographic processing, it is expected that the methodology can be used to fabricate label-free DNA microarrays and may be applicable to tissue engineering, which aims to create living tissue substitutes from cells seeded onto 3D scaffolds. Figure 1
Schematic illustration of fabrication procedures of SiNWs patterns 相似文献
16.
Du H Strohsahl CM Camera J Miller BL Krauss TD 《Journal of the American Chemical Society》2005,127(21):7932-7940
The separate developments of microarray patterning of DNA oligonucleotides, and of DNA hairpins as sensitive probes for oligonucleotide identification in solution, have had a tremendous impact on basic biological research and clinical applications. We have combined these two approaches to develop arrayable and label-free biological sensors based on fluorescence unquenching of DNA hairpins immobilized on metal surfaces. The thermodynamic and kinetic response of these sensors, and the factors important in hybridization efficiency, were investigated. Hybridization efficiency was found to be sensitive to hairpin secondary structure, as well as to the surface distribution of DNA hairpins on the substrate. The identity of the bases used in the hairpin stem as well as the overall loop length significantly affected sensitivity and selectivity. Surface-immobilized hairpins discriminated between two sequences with a single base-pair mismatch with high sensitivity (over an order of magnitude difference in signal) under identical assay conditions (no change in stringency). This represents a significant improvement over other microarray-based techniques. 相似文献
17.
Simon Q Lud Michael G Nikolaides Ilka Haase Markus Fischer Andreas R Bausch 《Chemphyschem》2006,7(2):379-384
For many biotechnological applications the label-free detection of biomolecular interactions is becoming of outstanding importance. In this Article we report the direct electrical detection of small peptides and proteins by their intrinsic charges using a biofunctionalized thin-film resistor. The label-free selective and quantitative detection of small peptides and proteins is achieved using hydrophobized silicon-on-insulator (SOI) substrates functionalized with lipid membranes that incorporate metal-chelating lipids. The response of the nanometer-thin conducting silicon film to electrolyte screening effects is taken into account to determine quantitatively the charges of peptides. It is even possible to detect peptides with a single charge and to distinguish single charge variations of the analytes even in physiological electrolyte solutions. As the device is based on standard semiconductor technologies, parallelization and miniaturization of the SOI-based biosensor is achievable by standard CMOS technologies and thus a promising basis for high-throughput screening or biotechnological applications. 相似文献
18.
Chang MJ Pang CR Liu J Bai H Deng J Xu ZG Zhang HL 《Journal of colloid and interface science》2011,360(2):826-833
High spatial resolution and large area thickness mapping of label-free protein microarray has been achieved using imaging ellipsometry (IE) under optimized conditions. The protein patterns with feature size down to 8×8 μm(2) was readily imaged, and the binding between the surface immobilized antigen and the antibody was monitored. Quantitative thickness analysis of antibody-antigen binding on the 32×32 μm(2) micron spots was successfully performed, and we have obtained a limit of detection as low as 1.2 pg/spot. This work demonstrates that appropriately optimized IE could be used as a highly sensitive and high through-put label-free technique for studying surface antigen-antibody recognition in sub-40 μm scale. 相似文献
19.
Gruber K Hermann BA Seeberger PH 《Angewandte Chemie (International ed. in English)》2011,50(37):A46-A51
Carbohydrates are important mediators of many biological processes that underlie cellular communication and disease mechanisms. Therapeutic agents include carbohydrate-based vaccines and the potent anti-viral protein Cyanovirin-N (CV-N). CV-N acts by specifically binding the carbohydrate structures decorating the cell surface of deadly viruses including human immunodeficiency virus (HI-V) or Ebola. In search for new carbohydrate-binding proteins and the development of sensors that exploit carbohydrate-protein interactions the label-free cantilever array technique can provides a fast, parallel and low-cost approach. 相似文献
20.
Cell-based impedance biosensing is an emerging technology that can be used to non-invasively and instantaneously detect and analyze cell responses to chemical and biological agents. This article highlights the fabrication and measurement technologies of cell impedance sensors, and their application in toxin detection and anti-cancer drug screening. We start with an introduction that describes the capability and advantages of cell-based sensors over conventional sensing technology, followed by a discussion of the influence of cell adhesion, spreading and viability during cell patterning on the subsequent impedance measurements and sensing applications. We then present an electronic circuit that models the cell-electrode system, by which the cellular changes can be detected in terms of impedance changes of the circuit. Finally, we discuss the current status on using cell impedance sensors for toxin detection and anti-cancer drug screening. 相似文献