Planar Optical Waveguide Based Sandwich Assay Sensors and Processes for the Detection of Biological Targets Including Early Detection of Cancers

Planar Optical Waveguide Based Sandwich Assay Sensors and Processes for the Detection of Biological Targets Including Early Detection of Cancers
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Release: 2009
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An assay element is described including recognition ligands adapted for binding to carcinoembryonic antigen (CEA) bound to a film on a single mode planar optical waveguide, the film from the group of a membrane, a polymerized bilayer membrane, and a self-assembled monolayer containing polyethylene glycol or polypropylene glycol groups therein and an assay process for detecting the presence of CEA is described including injecting a possible CEA-containing sample into a sensor cell including the assay element, maintaining the sample within the sensor cell for time sufficient for binding to occur between CEA present within the sample and the recognition ligands, injecting a solution including a reporter ligand into the sensor cell; and, interrogating the sample within the sensor cell with excitation light from the waveguide, the excitation light provided by an evanescent field of the single mode penetrating into the biological target-containing sample to a distance of less than about 200 nanometers from the waveguide thereby exciting any bound reporter ligand within a distance of less than about 200 nanometers from the waveguide and resulting in a detectable signal.

Planar Optical Waveguide Based Sandwich Assay Sensors and Processes for the Detection of Biological Targets Including Protein Markers, Pathogens and Cellular Debris

Planar Optical Waveguide Based Sandwich Assay Sensors and Processes for the Detection of Biological Targets Including Protein Markers, Pathogens and Cellular Debris
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Release: 2009
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An assay element is described including recognition ligands bound to a film on a single mode planar optical waveguide, the film from the group of a membrane, a polymerized bilayer membrane, and a self-assembled monolayer containing polyethylene glycol or polypropylene glycol groups therein and an assay process for detecting the presence of a biological target is described including injecting a biological target-containing sample into a sensor cell including the assay element, with the recognition ligands adapted for binding to selected biological targets, maintaining the sample within the sensor cell for time sufficient for binding to occur between selected biological targets within the sample and the recognition ligands, injecting a solution including a reporter ligand into the sensor cell; and, interrogating the sample within the sensor cell with excitation light from the waveguide, the excitation light provided by an evanescent field of the single mode penetrating into the biological target-containing sample to a distance of less than about 200 nanometers from the waveguide thereby exciting the fluorescent-label in any bound reporter ligand within a distance of less than about 200 nanometers from the waveguide and resulting in a detectable signal.

Toward Photostable Multiplex Analyte Detection on a Single Mode Planar Optical Waveguide

Toward Photostable Multiplex Analyte Detection on a Single Mode Planar Optical Waveguide
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Release: 2009
Genre:
ISBN:

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We have developed a waveguide-based optical biosensor for the sensitive and specific detection of biomarkers associated with disease. Our technology combines the superior optical properties of single-mode planar waveguides, the robust nature of functionalized self-assembled monolayer sensing films and the specificity of fluorescence sandwich immunoassays to detect biomarkers in complex biological samples such as serum, urine and sputum. We have previously reported the adaptation of our technology to the detection of biomarkers associated with breast cancer and anthrax. However, these approaches primarily used phospholipid bilayers as the functional film and organic dyes (ex: AlexaFluors) as the fluorescence reporter. Organic dyes are easily photodegraded and are not amenable to multiplexing because of their narrow Stokes' shift. Here we have developed strategies for conjugation of the detector antibodies with quantum dots for use in a multiplex detection platform. We have previously evaluated dihydroxylipoic acid quantum dots for the detection of a breast cancer biomarker. In this manuscript, we investigate the detection of the Bacillus anthracis protective antigen using antibodies conjugated with polymer-coated quantum dots. Kinetics of binding on the waveguide-based biosensor is reported. We compare the sensitivity of quantum dot labeled antibodies to those labeled with AlexaFluor and demonstrate the photostability of the former in our assay platform. In addition, we compare sulfydryl labeling of the antibody in the hinge region to that of nonspecific amine labeling. This is but the first step in developing a multiplex assay for such biomarkers on our waveguide platform.

Multichannel Waveguides for the Simultaneous Detection of Disease Biomarkers

Multichannel Waveguides for the Simultaneous Detection of Disease Biomarkers
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Release: 2009
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The sensor team at the Los Alamos National Laboratory has developed a waveguide-based optical biosensor that has previously been used for the detection of biomarkers associated with diseases such as tuberculosis, breast cancer, anthrax and influenza in complex biological samples (e.g., serum and urine). However, no single biomarker can accurately predict disease. To address this issue, we developed a multiplex assay for the detection of components of the Bacillus anthracis lethal toxin on single mode planar optical waveguides with tunable quantum dots as the fluorescence reporter. This limited ability to multiplex is still insufficient for accurate detection of disease or for monitoring prognosis. In this manuscript, we demonstrate for the first time, the design, fabrication and successful evaluation of a multichannel planar optical waveguide for the simultaneous detection of at least three unknown samples in quadruplicate. We demonstrate the simultaneous, rapid (30 min), quantitative (with internal standard) and sensitive (limit of detection of 1 pM) detection of protective antigen and lethal factor of Bacillus anthracis in complex biological samples (serum) using specific monoclonal antibodies labeled with quantum dots as the fluorescence reporter.

Optimizing a Waveguide-based Sandwich Immunoassay for Tumor Biomarkers

Optimizing a Waveguide-based Sandwich Immunoassay for Tumor Biomarkers
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Release: 2008
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The sensor team at the Los Alamos National Laboratory has developed a waveguide-based optical biosensor for the detection of biomarkers associated with the disease. We have previously demonstrated the application of this technology to the sensitive detection of carcinoembryonic antigen in serum and nipple aspirate fluid from breast cancer patients. In this publication, we report improvements to this technology that will facilitate transition to a point-of-care diagnostic system and/or robust research tool.

Optical Biosensors: Present & Future

Optical Biosensors: Present & Future
Author: Frances S. Ligler
Publisher: Gulf Professional Publishing
Total Pages: 626
Release: 2002-04-16
Genre: Science
ISBN: 9780444509741

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PART I. Optical Biosensors: The Present -- Chapter 1. Optrode-based Fiber Optic Biosensors -- Israel Biran and David R. Walt -- Chapter 2. Evanescent Wave Fiber Optic Biosensors -- Chris Rowe Taitt and Frances S. Ligler -- Chapter 3. Planar Waveguides for Fluorescence Biosensors -- Kim Sapsford, Chris Rowe Taitt, and Frances S. Ligler -- Chapter 4. Flow Immnosensor -- Anne W. Kusterbeck -- Chapter 5. Time Resolved Fluorescence -- Richard Thompson -- Chapter 6. Electrochemiluminescence -- Mark M. Richter -- Chapter 7. Surface Plasmon Resonance Biosensors -- Jiri Homola, Sinclair Yee, and David Myszka -- Chapter 8. The Resonant Mirror Optical Biosensor -- Tim Kinning and Paul Edwards -- Chapter 9. Interferometric Biosensors -- Daniel P. Campbell and Candice J. McCloskey -- Part II. Optical Biosensors: The Future -- Chapter 10. Genetic Engineering of Signaling Molecules -- Agatha Feltus and Sylvia Daunert -- Chapter 11. Artificial Receptors for Chemosensors -- Thomas W. Bell and Nicholas ...

Towards Fieldable Rapid Bioagent Detection: Advanced Resonant Optical Waveguide and Biolayer Structures for Integrated Biosensing

Towards Fieldable Rapid Bioagent Detection: Advanced Resonant Optical Waveguide and Biolayer Structures for Integrated Biosensing
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Total Pages: 23
Release: 2007
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ISBN:

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This research project established the understanding necessary for the stacked planar affinity-regulated resonate optical waveguide (SPARROW) architecture to serve as a transducer device platform for rapid biomolecular detection. SPARROW structures have been grown with waveguide losses of less than 1 dB/cm. Thin biolayers of less than 10 nm were achieved and SAM attachment to polycrystalline surfaces studied. SPARROW transducer measurements were conducted using sucrose analyte solutions applied to the top waveguide surface via a PDMS microfluidic channel. For a flow channel and waveguide interaction width of 1200 microns, detection equivalent to a surface loading of from 10 to 1 pg/mm^2 was achieved under different SPARROW surface conditions. Achievable detection was modeled and indicated a Limit of Detection (LODs) lower than 0.1 pg/mm^2 for spore sized targets.

Planar Total Internal Reflection Biofouling Sensors

Planar Total Internal Reflection Biofouling Sensors
Author: Koo Hyun Nam
Publisher:
Total Pages: 154
Release: 2010
Genre:
ISBN:

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Planar, integrated microscale sensors utilizing prism-coupler type angular interrogation sensing technique have been demonstrated. The main structure of the sensor consists of an optical prism coupled to a built-in waveguide to introduce Fraunhofer diffraction when light ray comes into the prism from the waveguide. The Fraunhofer diffraction creates spectrum of consecutive rays over the sensing edge of the prism such that there is no need for the bulky scanning mechanisms typically used in other macro scale sensing systems. Two types of sensors are presented: (1) total internal reflection based critical point detection (CPD) sensor, and (2) surface plasmon resonance (SPR) based resonance point detection (RPD) sensor. The CPD sensor is fabricated by a simple, two-mask process which creates a right angle prism with three sides with lengths of 1, 0.86, and 1.33 mm, respectively in the prototype design and a waveguide with a cross sectional area of 4~0.25 um2. The 0.25 um-thick core and the 2.5 um-thick cladding layers of the waveguide are made of silicon nitride and silicon dioxide, respectively. The CPD sensing technique measures the shift of the critical point of the total reflection as the results of change of refractive index due to biofouling. Optical simulations are used to validate the working principle and the calculated biofouling sensitivity is comparable to the other optical sensing methods. A baseline measurement has been conducted to verify the operation of the sensor with an error of less than ± 0.002 R.I.U. During a 9-hour biofouling measurement using milk as the media, a change in the refractive index as much as 0.0089 is recorded as the result of biofouling. The RPD sensing technique employs surface plasmon resonance as its sensing mechanism by measuring the shift of the resonance point with respect to the change of the incident angle. The design and fabrication process is similar to the fundamental structure of CPD sensors with an additional deposition of a thin metal layer on the sensing edge of the prism. The theoretical sensitivity is calculated as 90 deg RIU-1, which is comparable with the state-of-the-art optical sensors at 127 deg RIU-1. The refractive index measurement for selected liquids agrees with the values in the literature with an error range of less than ± 0.002 R.I.U. Furthermore, the refractive index change of biofouling formation is measured to be 0.0078 for a 9-hour experiment using milk as the testing media.

Enzyme-linked Immunosorbent Assay (ELISA)

Enzyme-linked Immunosorbent Assay (ELISA)
Author: Samira Hosseini
Publisher: Springer
Total Pages: 124
Release: 2017-12-30
Genre: Technology & Engineering
ISBN: 981106766X

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This book offers comprehensive information on all aspects of ELISA, starting with the fundamentals of the immune system. It also reviews the history of analytical assays prior to the advent of ELISA (enzyme-linked immunosorbent assay) and addresses the materials of choice for the fabrication of the platforms, possible biomolecular interactions, different protocols, and evaluation parameters. The book guides readers through the respective steps of the analytical assay, while also familiarizing them with the possible sources of error in the assay. It offers detailed insights into the immobilization techniques used for protein attachment, as well as methods for evaluating the assay and calculating the key parameters, such as sensitivity, specificity, accuracy and limit of detection. In addition, the book explores the advantages and shortcomings of the conventional ELISA, as well as various approaches to improving its performance. In this regard, merging and integrating other technologies with widely known ELISAs have opened new avenues for the advancement of this immunoassay. Accordingly, the book provides cutting-edge information on integrated platforms such as ELISpot, plasmonic ELISAs, sphere-/bead-based ELISAs, paper-/fiber-based ELISAs and ELISA in micro-devices.

Metal Nanoclusters in Catalysis and Materials Science: The Issue of Size Control

Metal Nanoclusters in Catalysis and Materials Science: The Issue of Size Control
Author: Benedetto Corain
Publisher: Elsevier
Total Pages: 471
Release: 2011-08-11
Genre: Technology & Engineering
ISBN: 0080555004

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Metal Nanoclusters in Catalysis and Materials Science: The Issue of Size Control deals with the synthesis of metal nanoclusters along all known methodologies. Physical and chemical properties of metal nanoclusters relevant to their applications in chemical processing and materials science are covered thoroughly. Special attention is given to the role of metal nanoclusters size and shape in catalytic processes and catalytic applications relevant to industrial chemical processing.An excellent text for expanding the knowledge on the chemistry and physics of metal nanoclusters. Divided in two parts; Part I deals with general aspects of the matter and Part II has to be considered a useful handbook dealing with the production of metal nanoclusters, especially from their size-control point of view. * Divided into two parts for ease of reference: general and operational * Separation of synthetic aspects, physical properties and applications* Specific attention is given to the task of metal nanoclusters size-control