Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

FRETS-VWF73, a first fluorogenic substrate for ADAMTS13 assay.

Koichi Kokame1, Yuko Nobe, Yoshihiro Kokubo

  • 1National Cardiovascular Centre Research Institute, Suita, Osaka, Japan. kame@ri.ncvc.go.jp

British Journal of Haematology
|April 2, 2005
PubMed
Summary

A new assay rapidly measures ADAMTS13 enzyme activity, crucial for diagnosing thrombotic thrombocytopenic purpura (TTP). This fluorescence-based method offers a faster, more accessible diagnostic tool for TTP and related thrombotic diseases.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Nonlinear Associations Between Sleep Duration and Incident LTCI Certification in Older Adults: Findings From the Iwate-Kenpoku Cohort Study.

Journal of the American Medical Directors Association·2026
Same author

Autofluorescence enables detection and quantification of EETosis in human leukocyte cultures.

Journal of leukocyte biology·2026
Same author

Oral Health and Cardiovascular Disease Among Hypertensive Medicated Adults With Uncontrolled Blood Pressure.

Journal of clinical hypertension (Greenwich, Conn.)·2026
Same author

Association between obesity and medical expenditures among Japanese adults treated for diabetes: A secondary analysis.

PloS one·2026
Same author

A long first intron of PROS1 modulates protein S expression across tissues in mice and humans.

Blood advances·2026
Same author

Long-term cardiovascular mortality risk of hyperuricemia and chronic kidney disease in a general Japanese population: NIPPON DATA90.

Endocrine·2026

Area of Science:

  • Biochemistry
  • Hematology
  • Enzymology

Background:

  • ADAMTS13 is a plasma metalloprotease that regulates von Willebrand factor (VWF) activity.
  • Deficiency in ADAMTS13 activity causes thrombotic thrombocytopenic purpura (TTP) due to VWF multimer accumulation.
  • Current methods for measuring ADAMTS13 activity are time-consuming and require specialized expertise.

Purpose of the Study:

  • To develop a rapid and efficient assay for quantifying ADAMTS13 activity.
  • To establish a fluorescence resonance energy transfer (FRET) based assay using a synthetic substrate.
  • To enable faster diagnosis and characterization of thrombotic microangiopathies.

Main Methods:

  • Development of a synthetic peptide substrate, FRETS-VWF73, containing modified residues.

Related Experiment Videos

  • Utilizing FRET technology where cleavage of the substrate by ADAMTS13 relieves fluorescence quenching.
  • Incubation of the substrate with human plasma samples and measurement of fluorescence in a 96-well plate format.

    • Main Results:

    • The FRETS-VWF73 assay demonstrated quantitative measurement of ADAMTS13 activity within one hour.
    • Normal human plasma showed increased fluorescence over time, indicating substrate cleavage.
    • ADAMTS13-deficient plasma did not result in increased fluorescence, confirming assay specificity.
    • The assay is compatible with standard laboratory equipment, including commercial plate readers.

    Conclusions:

    • The developed FRET assay provides a rapid, quantitative, and accessible method for measuring ADAMTS13 activity.
    • This assay can aid in the diagnosis and management of thrombotic thrombocytopenic purpura (TTP).
    • The assay holds potential for predicting thrombotic disease risk by assessing ADAMTS13 activity levels.