Mass-Sensing Multianalyte Micro-Array Based Immunoassay by Direct Nir Fluorescence and Quantitative Image Analysis

Silzel, John W.; Dodson, Charles; Obremski, Robert J.; Tsay, Tsong

doi:10.1007/978-94-011-5286-0_15

Cited by 18 publications

(19 citation statements)

References 4 publications

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“…Microarrays of antibodies (14,15) or other binding molecules, such as avidin (16), can be "printed" onto plastic sheets or into assay plates using inkjet microspotting technology, and ligand binding can be interrogated with a fluorescence microscope, a charge-coupled device camera, or an imaging instrument like the MVF. While the technology appears to work successfully for immunoassays (14,16) or nucleic acid hybridization assays (17), adaptation to ligand-receptor binding assays is likely to present significant challenges. The manufacturing process for these arrays is complex, and receptors are more likely than antibodies to lose activity when they are dried onto a surface.…”

Section: Discussionmentioning

confidence: 98%

A Generic Particle-Based Nonradioactive Homogeneous Multiplex Method for High-Throughput Screening Using Microvolume Fluorimetry

Martens

Bakker

Rodríguez

et al. 1999

Analytical Biochemistry

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Section: Discussionmentioning

confidence: 98%

A Generic Particle-Based Nonradioactive Homogeneous Multiplex Method for High-Throughput Screening Using Microvolume Fluorimetry

Martens

Bakker

Rodríguez

et al. 1999

Analytical Biochemistry

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“…The ELF-based assay required a specially constructed CCD-based camera to read the signal, and a sensitivity of approximately 10 ng͞ml was achieved. In another report, Abs labeled with a near-infrared dye were used to detect IgG subclasses on a microarray (18); this system also required a specially designed imaging system and achieved a detection limit of approximately 15 ng͞ml. In contrast, signal amplification by immunoRCA gives sensitivity in the pg͞ml range and allows the assay results to be read with commonly available microarray scanners.…”

Section: Discussionmentioning

confidence: 99%

Immunoassays with rolling circle DNA amplification: A versatile platform for ultrasensitive antigen detection

Schweitzer

Wiltshire

Lambert

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

536

390

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We describe an adaptation of the rolling circle amplification (RCA) reporter system for the detection of protein Ags, termed ''immunoRCA.'' In immunoRCA, an oligonucleotide primer is covalently attached to an Ab; thus, in the presence of circular DNA, DNA polymerase, and nucleotides, amplification results in a long DNA molecule containing hundreds of copies of the circular DNA sequence that remain attached to the Ab and that can be detected in a variety of ways. Using immunoRCA, analytes were detected at sensitivities exceeding those of conventional enzyme immunoassays in ELISA and microparticle formats. The signal amplification afforded by immunoRCA also enabled immunoassays to be carried out in microspot and microarray formats with exquisite sensitivity. When Ags are present at concentrations down to fM levels, specifically bound Abs can be scored by counting discrete fluorescent signals arising from individual Ag-Ab complexes. Multiplex immunoRCA also was demonstrated by accurately quantifying Ags mixed in different ratios in a two-color, single-molecule-counting assay on a glass slide. ImmunoRCA thus combines high sensitivity and a very wide dynamic range with an unprecedented capability for single molecule detection. This Ag-detection method is of general applicability and is extendable to multiplexed immunoassays that employ a battery of different Abs, each labeled with a unique oligonucleotide primer, that can be discriminated by a color-coded visualization system. ImmunoRCA-profiling based on the simultaneous quantitation of multiple Ags should expand the power of immunoassays by exploiting the increased information content of ratio-based expression analysis.prostate-specific antigen ͉ human IgE ͉ IgG ͉ ELISA ͉ immuno-microarrays

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“…By reducing the well volume and increasing the number of wells per plate, a significant improvement in the performance of microtiter plate-based methods has been achieved [1][2][3]. Another approach, inspired by the success of DNA chips [4], employs small protein microarrays deposited upon various substrates [5][6][7][8]. Such arrays are suitable for multianalyte analysis but in most cases are limited to a single sample for each run.…”

Section: Introductionmentioning

confidence: 99%

Multichannel homogeneous immunoassay for detection of 2,4,6‐trinitrotoluene (TNT) using a microfabricated capillary array electrophoresis chip

Bromberg

Mathies

2004

Electrophoresis

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A high-throughput homogeneous immunoassay for the sensitive detection of 2,4,6-trinitrotoluene (TNT) has been developed using radial capillary array electrophoresis microdevices. Samples consisting of equilibrium mixtures of anti-TNT antibody (Ab), fluorescein-labeled TNT, and various concentrations of unlabeled TNT were electrokinetically injected into 48 channels of a radial capillary array electrophoresis microchannel plate. The rapid electrophoretic separation allows us to analyze the equilibrium ratio formed by the competition between the labeled and the unlabeled TNT for Ab binding. The simultaneous parallel TNT separations facilitate determination of a calibration curve for the TNT assay, which has high sensitivity (LOD, 1 ng/mL) and a wide dynamic range (1-300 ng/mL).

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Mass-Sensing Multianalyte Micro-Array Based Immunoassay by Direct Nir Fluorescence and Quantitative Image Analysis

Cited by 18 publications

References 4 publications

A Generic Particle-Based Nonradioactive Homogeneous Multiplex Method for High-Throughput Screening Using Microvolume Fluorimetry

A Generic Particle-Based Nonradioactive Homogeneous Multiplex Method for High-Throughput Screening Using Microvolume Fluorimetry

Immunoassays with rolling circle DNA amplification: A versatile platform for ultrasensitive antigen detection

Multichannel homogeneous immunoassay for detection of 2,4,6‐trinitrotoluene (TNT) using a microfabricated capillary array electrophoresis chip

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