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Related Experiment Videos

A sensitive competitive binding assay for exogenous and endogenous heparins

J Dawes, D S Pepper

    Thrombosis Research
    |August 15, 1982
    PubMed
    Summary

    A novel assay accurately measures heparin levels using competitive binding to protamine-Sepharose. This sensitive method quantifies heparin in biological samples, aiding research on heparin

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    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Pharmacology

    Background:

    • Heparins are crucial anticoagulants with complex structures.
    • Accurate quantification of heparins in biological samples is essential for research and clinical applications.
    • Existing methods may lack sensitivity or specificity for endogenous heparin analysis.

    Purpose of the Study:

    • To develop a sensitive and versatile assay for heparin quantification.
    • To measure endogenous heparin concentrations in human biological fluids.

    Main Methods:

    • A competitive binding assay utilizing 125I-labelled heparin and protamine-Sepharose.
    • The assay measures heparin concentration based on its ability to compete for binding.
    • Sample pretreatment with protease was employed for analyzing biological fluids.

    Main Results:

    • The assay demonstrated high sensitivity, detecting heparin concentrations as low as 10 ng/ml.
    • Assay response correlated with the degree of sulfation and molecular weight of acidic polysaccharides.
    • Endogenous heparins were successfully quantified in normal human serum, plasma, and urine.

    Conclusions:

    • A novel, highly sensitive assay for heparin quantification has been successfully developed.
    • The assay is versatile and suitable for measuring endogenous and exogenous heparins in biological samples.
    • This method holds significant potential for advancing research in heparin-related studies.

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