A Review on Functionalized Gold Nanoparticles for Biosensing Applications

Nanoparticle technology plays a key role in providing opportunities and possibilities for the development of new generation of sensing tools. The targeted sensing of selective biomolecules using functionalized gold nanoparticles (Au NPs) has become a major research thrust in the last decade. Au NP-based sensors are expected to change the very foundations of sensing and detecting biomolecules. In this review, we will discuss the use of surface functionalized Au NPs for smart sensor fabrication leading to detection of specific biomolecules and heavy metal ions.     

High efficiency Hall effect micro-biosensor platform for detection of magnetically labeled biomolecules

Detection of magnetically labeled biomolecules using micro-Hall biosensors is a promising method for monitoring biomolecular recognition processes. The measurement efficiency of standard systems is limited by the time taken for magnetic beads to reach the sensing area of the Hall devices. Here, micro-current lines were integrated with Hall effect structures to manipulate the position of magnetic beads via field gradients generated by localized currents flowing in the current lines. Beads were accumulated onto the sensor surface within seconds of passing currents through the current lines. Real-time detection of magnetic beads using current lines integrated with Hall biosensors was achieved. These results are promising in establishing Hall biosensor platforms as efficient and inexpensive means of monitoring biomolecular reactions for medical applications.     

Surface plasmon resonance biosensors incorporating gold nanoparticles

SPR biosensing is increasingly popular for the detection of a multitude of biomolecules. It offers label-free detection and study of proteins, nucleic acids, and other biomolecules in real time. A recent trend involves incorporation of AuNPs, either within the sensing surface itself or as signal enhancing tagging molecules. The importance of AuNP and detecting agent spacing is described and techniques using macromolecular spacing aids are highlighted. Recent methods to enhance SPR detection capabilities using gold nanoparticles are reviewed, as well as device fabrication and the results of incorporation. SPR detection is a highly versatile method for the detection of biomolecules and, with the incorporation of AuNPs, shows promise in extending it to a number of new applications.     

Detection and manipulation of biomolecules by magnetic carriers

The detection and manipulation of single molecules on a common platform would be of great interest for basic research of biological or chemical systems. A promising approach is the application of magnetic carriers. The principles are demonstrated in this contribution. It is shown that paramagnetic beads can be detected by highly sensitive magnetoresistive sensors yielding a purely electronic signal. Different configurations are discussed. The capability of the sensors to detect even single markers is demonstrated by a model experiment. In addition, the paramagnetic beads can be used as carriers for biomolecules. They can be manipulated on-chip via currents running through specially designed line patterns. Thus, magnetic markers in combination with magnetoresistive sensors are a promising choice for future integrated lab-on-a-chip systems.     

A reagentless, disposable and multiplexed electronic biosensing platform: application to molecular logic gates

The construction of a reagentless, sensitive, disposable and multiplexed electronic sensing platform for one-spot simultaneous determination of biomolecules with significant difference in size (proteins and small molecules) is described. The sensing surface is fabricated by the hybridization of two types of redox-tags conjugated aptamers with the corresponding complementary DNAs, which are self-assembled on a gold nanoparticle-modified screen printed carbon electrode. The presence of the target analytes leads to the release of the tagged signaling aptamers from the sensing surface, and the surface-remained tags exhibit well-resolved peaks, whose positions and sizes reflect the identities and concentrations of the target analytes, respectively. The application of the proposed sensing platform for molecular logic gate operations is also demonstrated.     

Nanobiohybrid Material‐Based Bioelectronic Devices

Biomolecules, especially proteins and nucleic acids, have been widely studied to develop biochips for various applications in scientific fields ranging from bioelectronics to stem cell research. However, restrictions exist due to the inherent characteristics of biomolecules, such as instability and the constraint of granting the functionality to the biochip. Introduction of functional nanomaterials, recently being researched and developed, to biomolecules have been widely researched to develop the nanobiohybrid materials because such materials have the potential to enhance and extend the function of biomolecules on a biochip. The potential for applying nanobiohybrid materials is especially high in the field of bioelectronics. Research in bioelectronics is aimed at realizing electronic functions using the inherent properties of biomolecules. To achieve this, various biomolecules possessing unique properties have been combined with novel nanomaterials to develop bioelectronic devices such as highly sensitive electrochemical‐based bioelectronic sensing platforms, logic gates, and biocomputing systems. In this review, recently reported bioelectronic devices based on nanobiohybrid materials are discussed. The authors believe that this review will suggest innovative and creative directions to develop the next generation of multifunctional bioelectronic devices.     

Atomic force microscopy-based cancer diagnosis by detecting cancer-specific biomolecules and cells

Atomic force microscopy (AFM) has recently attracted much attention due to its ability to analyze biomolecular interactions and to detect certain biomolecules, which play a crucial role in disease expression. Despite recent studies reporting AFM imaging for the analyses of biomolecules, the application of AFM-based cancer-specific biomolecule/cell detection has remained largely underexplored, especially for the early diagnosis of cancer. In this paper, we review the recent attempts, including our efforts, to analyze and detect cancer-specific biomolecules and cancer cells. We particularly focus on two AFM-based cancer diagnosis techniques: (i) AFM imaging-based biomolecular and cellular detection, (ii) AFM cantilever-based biomolecular sensing and cell analysis. It is shown that AFM-based biomolecular detection has been applied for not only early diagnosing cancer, by measuring the minute amount of cancer-specific proteins, but also monitoring of cancer progression, by correlating the amount of cancer-specific proteins with the progression of cancer. In addition, AFM-based cell imaging and detection have been employed for diagnosing cancer, by detecting cancerous cells in tissue, as well as understanding cancer progression, by characterizing the dynamics of cancer cells. This review, therefore, highlights AFM-based biomolecule/cell detection, which will pave the way for developing a fast and point-of-care diagnostic system for biomedical applications.     

TiO2 nanowire FET device: encapsulation of biomolecules by electro polymerized pyrrole propylic acid

Silane-based methods have become the standards for the conjugation of biomolecules, especially for the preparation of one-dimensional nanomaterial biosensors. However, the specific binding of those target molecules might raise problems with regard to the sensing and non-sensing regions, which may contaminate the sensing devices and decrease their sensitivity. This paper attempts to explore the encapsulation of biomolecules on a one-dimensional nanomaterial field effect transistor (FET) biosensor using polypyrrole propylic acid (PPa). Specifically, the encapsulation of biomolecules via the electropolymerization of pyrrole propylic acid (Pa), a self-made low-conductivity polymer, on TiO(2)-nanowire (NW)-based FETs is presented. The energy dispersive spectrum (EDS) was obtained and electrical analysis was conducted to investigate PPa entrapping anti-rabbit IgG (PPa/1°Ab) on a composite film. The specificity, selectivity and sensitivity of the sensor were analyzed in order to determine the immunoreaction of PPa/1°Ab immobilized NW biosensors. Our results show that PPa/1°Ab achieved high specificity immobilization on NWs under the EDS analysis. Furthermore, the TiO(2)-NW FET immunosensor developed in this work successfully achieved specificity, selectivity and sensitivity detection for the target protein rabbit IgG at the nano-gram level. The combination of PPa material and the electropolymerization method may provide an alternative method to immobilize biomolecules on a specific surface, such as NWs.     

Nanowire sensors for multiplexed detection of biomolecules

Nanowire-based detection strategies provide promising new routes to bioanalysis that could one day revolutionize the healthcare industry. This review covers recent developments in nanowire sensors for multiplexed detection of biomolecules such as nucleic acids and proteins. We focus on encoded nanowire suspension arrays and semiconductor nanowire-based field-effect transistors. Nanowire assembly and integration with microchip technology is emphasized as a key step toward the ultimate goal of multiplexed detection at the point of care using portable, low power, electronic biosensor chips.     

Carbon nanotubes and glucose oxidase bionanocomposite bridged by ionic liquid-like unit: preparation and electrochemical properties

For their biocompatibility and potential bionanoelectronic applications, integration of carbon nanotubes (CNTs) with biomolecules such as redox enzyme is highly anticipated. Therein, CNTs are expected to act not only as an electron transfer promoter, but also as immobilizing substrate for biomolecules. In this report, a novel method for immobilization of biomolecules on CNTs was proposed based on ionic interaction, which is of universality and widespread use in biological system. As illustrated, glucose oxidase (GOD) and single-walled carbon nanotubes (SWNTs) were integrated into a unitary bionanocomposite by means of ionic liquid-like unit on functionalized SWNTs. The resulted bionanocomposite illustrated better redox response of immobilized GOD in comparison of that prepared by weak physical absorption without ionic interaction. As a potential application of concept, the electrochemical detection of glucose was exemplified based on this novel bionanocomposite.     

Dynamic mass spectrometry probe for electrospray ionization mass spectrometry monitoring of bioreactors for therapeutic cell manufacturing

Large-scale manufacturing of therapeutic cells requires bioreactor technologies with online feedback control enabled by monitoring of secreted biomolecular critical quality attributes (CQAs). Electrospray ionization mass spectrometry (ESI-MS) is a highly sensitive label-free method to detect and identify biomolecules, but requires extensive sample preparation before analysis, making online application of ESI-MS challenging. We present a microfabricated, monolithically integrated device capable of continuous sample collection, treatment, and direct infusion for ESI-MS detection of biomolecules in high-salt solutions. The dynamic mass spectrometry probe (DMSP) uses a microfluidic mass exchanger to rapidly condition samples for online MS analysis by removing interfering salts, while concurrently introducing MS signal enhancers to the sample for sensitive biomolecular detection. Exploiting this active conditioning capability increases MS signal intensity and signal-to-noise ratio. As a result, sensitivity for low-concentration biomolecules is significantly improved, and multiple proteins can be detected from chemically complex samples. Thus, the DMSP has significant potential to serve as an enabling portion of a novel analytical tool for discovery and monitoring of CQAs relevant to therapeutic cell manufacturing.     

Gold nanoparticle-based signal amplification for biosensing

Colloidal gold nanoparticles (AuNPs), with unique properties such as highly resonant particle plasmons, direct visualization of single nanoclusters by scattering of light, catalytic size enhancement by silver deposition, conductivity, and electrochemical properties, are very attractive materials for several applications in biotechnology. Furthermore, as excellent biological tags, AuNPs can be easily conjugated with biomolecules and retain the biochemical activity of the tagged biomolecules, making AuNPs ideal transducers for several biorecognition applications. The goal of this article is to review recent advances of using AuNPs as labels for signal amplification in biosensing applications. We focus on the signal amplification strategies of AuNPs in biosensing/biorecognition, more specifically, on the main optical and electrochemical detection methods that involve AuNP-based biosensing. Particular attention is given to recent advances and trends in sensing applications.     

Analysis of native proteins from biological fluids by biomolecular interaction analysis mass spectrometry (BIA/MS): exploring the limit of detection, identification of non-specific binding and detection of multi-protein complexes

Biomolecular interaction analysis mass spectrometry (BIA/MS) is a two-dimensional analytical technique that quantitatively and qualitatively detects analytes of interests. In the first dimension, surface plasmon resonance (SPR) is utilized for detection of biomolecules in their native environment. Because SPR detection is non-destructive, analyte(s) retained on the SPR-active sensor surface can be analyzed in a second dimension using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The qualitative nature of the MALDI-TOF MS analysis complements the quantitative character of SPR sensing and overcomes the shortcomings of the SPR detection stemming from the inability to differentiate and characterize multi-protein complexes and non-specific binding. In this work, the benefit of performing MS analysis following SPR sensing is established. Retrieval and detection of four markers present in biological fluids (cystatin C, beta-2-microglobulin, urinary protein 1 and retinol binding protein) was explored to demonstrate the effectiveness of BIA/MS in simultaneous detection of clinically related biomarkers and delineation of non-specific binding. Furthermore, the BIA/MS limit of detection at very low SPR responses was investigated. Finally, detection of in-vivo assembled protein complexes was achieved for the first time using BIA/MS.     

Covalent bound sensing layers on surface acoustic wave (SAW) biosensors

This paper reports on the development of immunosensors based on commercially available surface acoustic wave (SAW) devices working at 380 MHz. Approaches for coating the sensor surface with a sensing layer of receptive biomolecules are presented and discussed. It was found that the sensitivity strongly relates to the immobilization method. Additionally, the sensitivity can be influenced by the density of accessible biomolecules on the active sensing area. Usually, by most of the standard immobilization procedures, two-dimensional layers of receptive biomolecules are obtained. We present a three-dimensional layer, which provides a higher absolute amount of recognition molecules. A dextran layer is photoimmobilized to the sensor surface and the recognition molecules are covalently embedded into the dextran matrix. The feasibility of specific immunosensing is investigated using SAW sensors connected to a fluid handling system.     

Fabrication of Carbon Nanotube High-Frequency Nanoelectronic Biosensor for Sensing in High Ionic Strength Solutions

The unique electronic properties and high surface-to-volume ratios of single-walled carbon nanotubes (SWNT) and semiconductor nanowires (NW) ^1-4 make them good candidates for high sensitivity biosensors. When a charged molecule binds to such a sensor surface, it alters the carrier density⁵ in the sensor, resulting in changes in its DC conductance. However, in an ionic solution a charged surface also attracts counter-ions from the solution, forming an electrical double layer (EDL). This EDL effectively screens off the charge, and in physiologically relevant conditions ~100 millimolar (mM), the characteristic charge screening length (Debye length) is less than a nanometer (nm). Thus, in high ionic strength solutions, charge based (DC) detection is fundamentally impeded^6-8.We overcome charge screening effects by detecting molecular dipoles rather than charges at high frequency, by operating carbon nanotube field effect transistors as high frequency mixers^9-11. At high frequencies, the AC drive force can no longer overcome the solution drag and the ions in solution do not have sufficient time to form the EDL. Further, frequency mixing technique allows us to operate at frequencies high enough to overcome ionic screening, and yet detect the sensing signals at lower frequencies^11-12. Also, the high transconductance of SWNT transistors provides an internal gain for the sensing signal, which obviates the need for external signal amplifier.Here, we describe the protocol to (a) fabricate SWNT transistors, (b) functionalize biomolecules to the nanotube¹³, (c) design and stamp a poly-dimethylsiloxane (PDMS) micro-fluidic chamber¹⁴ onto the device, and (d) carry out high frequency sensing in different ionic strength solutions¹¹.     

The nature of the charge carriers in solvated biomacromolecules

Solid state electrolysis experiments were performed on the biomolecules, hemoglobin, cytochromec, collagen, lecithin and melanin at various hydration states; and for hemoglobin at various solvation states with methanol adsorbate. The evolved hydrogen was measured and compared with theoretical (Faraday's Law) expectations for the known amount of charge passed through the adsorbents. The difference between the theoretical and actual is a measure of the contributions of electronic charge carriers to the total current. Thus the protonic/electronic conduction ratios are determined.All biomolecules tested appear to be mixed semiconductors. That is, both electronic and protonic charge carriers make significant contributions to the currents over hydration ranges from 6% to above 50%. The constant temperature conductivity increases exponentially with hydration (solvation) but the ratio of protonic to electronic conduction increases linearly with hydration for the globular proteins, hemoglobin and cytochromec. The fibrous protein, collagen, may be a protonic semiconductor in the dry state, with an electronic component that increases linearly with hydration. The hemoglobin-methanol system shows only electronic conductivity below 2 BET monolayers, with a sharp onset to 70% protonic conductivity above this value. This result is similar to the DNA-water system previously reported. The protonic/electronic ratio in hydrated hemoglobin may be a function of the applied voltage; being predominantly electronic below 30 volts (300 volts/cm), and a constant mixed value above 100 volts (1000 volts/cm). Our results suggest that both electronic and protonic conduction are intrinsic processes in these substances and subject to control by a number of techniques.     

Highly sensitive and selective detection of thiol-containing biomolecules using DNA-templated silver deposition

In this work, we report a new fluorescent method for the label-free assay of thiol-containing amino acids and peptide by use of silver deposited DNA duplex and the intercalating dye. The sensing approach is based on the specific interactions between thiols and DNA-templated silver deposition via robust Ag-S bonds. In the presence of thiols, the intercalating dye gives a dramatic increase in fluorescence as a result of the strong interaction between the intercalator and the released DNA from silver surfaces. The detection limit of this method is lower than or at least comparable to previous fluorescence-based methods, and the turn-on sensing mode offers additional advantage to efficiently reduce background noise. Moreover, the detection and discrimination process can be observed by the naked eye with the aid of an UV transilluminator. This method also exhibits excellent selectivity for these thiol-containing biomolecules over various other amino acids. As far as we know, our method is the first example using the specific interaction between thiols and silver-coated DNA to fabricate a turn-on fluorescent sensor for biological thiols with high sensitivity and selectivity.     

Two-dimensional PtSe2 Theoretically Enhanced Goos-Hänchen Shift Sensitive Plasmonic Biosensors

Platinum diselenide (PtSe₂), an emerging two-dimensional transition metal dichalcogenide, exhibits thickness-dependent refractive index, and hence, intriguing optical properties. Here, we employ it as a plasmonic sensing substrate to achieve significant enhancement in Goos-Hänchen shift sensitivity. Through systematic optimization of all parameters, four optimum sensing configurations have been achieved at different wavelengths ranging from visible to near-infrared region, where the Goos-Hänchen shift sensitivity receives four times enhancement in comparison with the conventional bare gold sensing substrate. There is a linear range of Goos-Hänchen shift with the tiny change of refractive index for each optimal configuration. The detection limit of the refractive index change can be as low as 5 × 10⁻⁷ RIU which is estimated to be lower by 2 orders of magnitude, and the corresponding sensitivity of biomolecules has a 1000-fold increment compared with that of bare gold-based sensors.

     

Integration of biomolecules and nanomaterials: towards highly selective and sensitive biosensors

Significant efforts have been made toward the development of high-performance biosensors for various applications. Advances in nanotechnology have resulted in the development of highly sensitive electrochemical sensing devices. It is believed that highly sensitive and selective biosensors can be realized through the integration of biomolecules and nanomaterial-based sensor platforms. Numerous articles have described combining biomolecules as recognition elements with nanotechnology for the development of biosensors with enhanced selectivity and sensitivity. Recent advances in the development of biosensors through the integration of biomolecules with nanotechnology are reviewed in this article.