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Enzyme-Linked Immunosorbent Assay01:33

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or...
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ELIME Enzyme Linked Immuno Magnetic Electrochemical Method for Mycotoxin Detection
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ADAMTS13 Activity Measurement by ELISA and Fluorescence Resonance Energy Transfer Assay.

Gary W Moore1,2,3, Marcos Llusa4, Margaret Griffiths4

  • 1Research and Development, Technoclone Herstellung von Diagnostika und Arzneimitteln GmbH, Vienna, Austria. gary.moore@addenbrookes.nhs.uk.

Methods in Molecular Biology (Clifton, N.J.)
|May 19, 2023
PubMed
Summary

Accurate ADAMTS13 activity measurement is key for diagnosing thrombotic thrombocytopenic purpura (TTP). Rapid enzyme-linked immunosorbent assays (ELISA) and fluorescence resonance energy transfer (FRET) provide timely results for effective patient management.

Keywords:
ADAMTS13ELISAFluorescence resonance energy transfer assayRapid assayThrombotic microangiopathyThrombotic thrombocytopenic purpura

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Area of Science:

  • Biochemistry
  • Hematology
  • Diagnostic Assays

Background:

  • Accurate ADAMTS13 activity measurement is critical for differentiating thrombotic thrombocytopenic purpura (TTP) from other thrombotic microangiopathies.
  • Traditional assays were time-consuming, delaying diagnosis and treatment decisions.
  • Rapid assays are now available to aid in acute diagnosis and management.

Purpose of the Study:

  • To describe the principles, performance, and practical aspects of ELISA and FRET assays for quantitative ADAMTS13 activity measurement.
  • To highlight the advantages of rapid assays in clinical settings.

Main Methods:

  • Describes enzyme-linked immunosorbent assays (ELISA) for ADAMTS13 activity.
  • Describes fluorescence resonance energy transfer (FRET) assays for ADAMTS13 activity.
  • Discusses practical considerations and performance of these quantitative assays.

Main Results:

  • ELISA assays provide results in approximately 4 hours without specialized equipment.
  • FRET and chemiluminescence assays yield results in under an hour but require specific analytical platforms.
  • These rapid assays enable timely diagnosis and management of TTP.

Conclusions:

  • Rapid ELISA and FRET assays significantly improve the diagnostic workflow for TTP.
  • Timely measurement of ADAMTS13 activity is essential for effective clinical management.
  • These assays offer practical solutions for laboratories in assessing ADAMTS13 activity.