Detection of biological threat agents by immunomagnetic microsphere-based solid phase fluorogenic- and electro-chemiluminescence.
Abstract: This article reviews the recent development of two solid-phase chemiluminescence-based techniques, fluorogenic-chemiluminescence (FCL) and electro-chemiluminescence (ECL) for detection of biological threat agents. Both techniques entail a labeled sandwich immunoassay. The objectives of this work are to develop advanced techniques for sensitive and effective detection of a target analyte, particularly in cases where the analysis includes complex samples containing multiple contaminating factors. Other important considerations in developing such detection techniques include the ease of use, the rapid determination of the results, and system automation for field applications. In FCL, alkaline phosphatase is used as a label and this technique utilizes the dual features of fluorescence and visual color generated upon the presence of the fluorogenic compound, AttoPhos. The assay reaction is determined by measuring the fluorescence. In ECL, the label is a ruthenium-trisbipyridal, which is excited to a higher energy state by an electric current-driven redox reaction, and the extent of the reaction is assessed via photon emission. Both techniques depend upon the magnetic separation technique as a means to isolate the target immunological agents from the sample for analysis. This magnetic capture system allows for a reaction to occur on the electron effective-transfer zone in the ECL and also provides the reaction site for the labeled sandwich in the FCL. Comparative studies of these two techniques for detection of biological threat agents have been performed and the advantages of using magnetic microspheres versus conventional solid-phase matrices are discussed.
Publication Date: 2000-08-17 PubMed ID: PMC10945457DOI: 10.1016/s0956-5663(99)00068-8Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Review
Summary
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The research article discusses the development and comparisons of two solid-phase chemiluminescence techniques, namely fluorogenic-chemiluminescence (FCL) and electro-chemiluminescence (ECL), aimed for the detection of biological threat agents.
Objective of the Research
- The study aims to develop advanced FCL and ECL techniques to efficiently and sensitively detect a target analyte in complex samples laden with multiple contaminants.
- Considerations for the development of these techniques include their ease of use, quick results delivery, and ability for system automation for field applications.
Fluorogenic-Chemiluminescence (FCL)
- In Fluorogenic-Chemiluminescence, an enzyme called alkaline phosphatase is utilized as a label.
- The distinguishing feature of FCL lies in its use of fluorescence and visual color upon the introduction of a fluorogenic compound, AttoPhos.
- In this method, the fluorescence measure determines the assay reaction.
- A magnetic capture system isolates the target immunological agents from the sample allowing the reaction to occur for the labeled sandwich within the FCL.
Electro-Chemiluminescence (ECL)
- In Electro-Chemiluminescence, a ruthenium-trisbipyridal serves as the label.
- Ruthenium-trisbipyridal is excited to a higher energy state through an electric current-driven redox reaction, with the reaction scale being assessed via the emission of photons or light particles.
- Similar to FCL, ECL also utilizes the magnetic separation technique to isolate target immunological agents from the sample, which facilitates a reaction on the electron effective-transfer zone.
Comparative Analysis
- The study performs comparative analysis between the FCL and ECL techniques in detecting biological threat agents.
- The research discusses the benefits of using magnetic microspheres over conventional solid-phase matrices.
Cite This Article
APA
Yu H, Raymonda JW, McMahon TM, Campagnari AA.
(2000).
Detection of biological threat agents by immunomagnetic microsphere-based solid phase fluorogenic- and electro-chemiluminescence.
Biosens Bioelectron, 14(10-11), 829-840.
https://doi.org/10.1016/s0956-5663(99)00068-8 Publication
Researcher Affiliations
- Chemical and Biological Defense Group, Veridian Engineering, Buffalo, NY 14225, USA. kent_yu@bio-rad.com
MeSH Terms
- Biological Warfare
- Biosensing Techniques
- Immunomagnetic Separation
- Luminescent Measurements
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