A Homogeneous and Multiplexed Immunoassay for High-Throughput Screening Using Fluorometric Microvolume Assay Technology

Swartzman, Elana; Miraglia, Sheri; Mellentin-Michelotti, Julia; Evangelista, Lolita; Yuan, Pau-Miau

doi:10.1006/abio.1999.4128

Cited by 113 publications

(75 citation statements)

References 10 publications

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“…Recently, several multiplex protein analysis technologies have been developed, including slide/microarray-based assays (Knight et al, 2004;Tam et al, 2002;Wiese et al, 2001) and flow cytometry-based assays (Fulton et al, 1997;Oliver et al, 1998;Swartzman et al, 1999;Cook et al, 2001). Flow cytometry-based systems are currently the most widely used multiplex biomarker analysis technology.…”

Section: Introductionmentioning

confidence: 99%

“…Flow cytometry-based systems are currently the most widely used multiplex biomarker analysis technology. The Luminex-100 system uses uniformly sized microspheres internally labeled with graded proportions of a red and a near infrared fluorophore, 658 and 712 nm, providing the capacity to interrogate and classify 100 discrete beads (Fulton et al, 1997;Oliver et al, 1998;Swartzman et al, 1999;Martins, 2002). This is in contrast to the BD CBA system that discriminates beads based upon fluorescence intensity from a single fluorophore, which limits the multiplexing capacity (Cook et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

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Validation and comparison of luminex multiplex cytokine analysis kits with ELISA: Determinations of a panel of nine cytokines in clinical sample culture supernatants

duPont¹,

Wang

Wadhwa

et al. 2005

Journal of Reproductive Immunology

272

181

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Problem: Analyses of the expression pattern of multiple cytokines are frequently required for characterization of the status of the immune system as it pertains to Th type bias and intrinsic levels of inflammation. Classically, analysis of cytokine expression patterns has been performed by enzymelinked immunosorbent assays (ELISA) for each separate analyte. A new technology, Luminex MAP ® , facilitates the simultaneous evaluation of multiple immune mediators with advantages of higher throughput, smaller sample volume, and lower cost. Validation of this technology has been limited to small sample sets, limited use of clinical study specimens, and use of non-commercial reagents. * Corresponding author. Methods: Ninety-six specimens from women over the course of their respective pregnancies were evaluated for cytokine concentrations using commercially available ELISA kits and commercially available Luminex MAP ® kits according to the manufacturers' directions. Correlations between data sets were evaluated using Pearson's correlation coefficient (r).Results: Excellent correlations were demonstrated for IL-1␤, IL-4, IL-5, IL-6, IL-10, IFN ␥, and TNF ␣, in contrast to IL-12 p70 and IL-13. Conclusions: Luminex multiplex technology has distinct advantages and is a valid alternative method to ELISA for the evaluation of the majority of cytokines tested and for the characterization of immune system status.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Validation and comparison of luminex multiplex cytokine analysis kits with ELISA: Determinations of a panel of nine cytokines in clinical sample culture supernatants

duPont¹,

Wang

Wadhwa

et al. 2005

Journal of Reproductive Immunology

272

181

View full text Add to dashboard Cite

show abstract

“…Labeled probe methods such as multilabel and spatially resolved assays can offer amplified detection signals for multianalyte immunoassays. Multilabel methods need several labels, such as radioisotopes (15 ), fluorescence dyes (16 ), enzymes (17,18 ), metal ions (12,14,19 ), or quantum dots (20,21 ), which limits their application. Spatially resolved array assays with a single label can perform simultaneous detection of analytes by both optical (22)(23)(24) and electrochemical (25 ) responses.…”

mentioning

confidence: 99%

A Disposable Multianalyte Electrochemical Immunosensor Array for Automated Simultaneous Determination of Tumor Markers

et al. 2007

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Background: Automated and convenient multianalyte detection with high throughput is increasingly needed in clinical diagnosis. We developed a disposable 4-by-2 array for programmed simultaneous amperometric immunoassay of 4 tumor markers. Methods: We used a screen-printed technique, 1-step immobilization method, and flow injection technique. We immobilized carcinoembryonic antigen, ␣-fetoprotein, ␤-human choriogonadotropin, and carcinoma antigen 125 as model analytes in a redox mediator-grafted, biopolymer-modified, screen-printed carbon electrode array to capture corresponding horseradish peroxidaselabeled antibodies in competitive immunoreactions. The simultaneous multianalyte immunoassay was automatically carried out to amperometrically monitor the mediator-catalyzed enzymatic response to hydrogen peroxide, which decreased in proportion to the concentrations of analytes in samples. Results: The multianalyte immunosensor array had a throughput of 60 samples/h and allowed simultaneous detection of carcinoembryonic antigen, ␣-fetoprotein, ␤-human choriogonadotropin, and carcinoma antigen 125 in clinical serum samples with concentrations up to 188 g/L, 250 g/L, 266 IU/L, and 334 kIU/L, respectively. The detection limits (limits of the blank, mean of blank plus 3 SD) were 1.1 g/L, 1.7 g/L, 1.2 IU/L, and 1.7 kIU/L. The inter-and intraassay imprecision (CVs) of the immunosensor arrays were <7.8% and <9.0%, respectively. The immunosensor arrays were stable for 28 days.

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“…In particular, IAC should be tested using rFabs encapsulated in a sol-gel glass (SGG) matrix. [74][75][76][77][78] When antibodies to additional PAHs become available, they should be tested in the newer, micro-scaled multiplex immunoassay [79][80][81] and microarray ELISA 82 formats.…”

Section: To What Extent Does the New Scientific Knowledge Bridge The mentioning

confidence: 99%

Engineered Antibodies for Monitoring of Polynuclear Aromatic Hydrocarbons

Karu¹,

Roberts²,

Li³

2002

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A Homogeneous and Multiplexed Immunoassay for High-Throughput Screening Using Fluorometric Microvolume Assay Technology

Cited by 113 publications

References 10 publications

Validation and comparison of luminex multiplex cytokine analysis kits with ELISA: Determinations of a panel of nine cytokines in clinical sample culture supernatants

Validation and comparison of luminex multiplex cytokine analysis kits with ELISA: Determinations of a panel of nine cytokines in clinical sample culture supernatants

A Disposable Multianalyte Electrochemical Immunosensor Array for Automated Simultaneous Determination of Tumor Markers

Engineered Antibodies for Monitoring of Polynuclear Aromatic Hydrocarbons

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