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A versatile microassay for elastase using succinylated elastin

S K Rao1, M Mathrubutham, A Karteron

  • 1Department of Surgery, Long Island Jewish Medical Center, Long Island Campus for Albert Einstein College of Medicine, New Hyde Park, New York 11042, USA. rao@lij.edu

Analytical Biochemistry
|August 1, 1997
PubMed
Summary
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A new elastase assay uses succinylated elastin to detect primary amines released by enzymatic digestion. This rapid, sensitive method quanties elastase in various biological samples.

Area of Science:

  • Biochemistry
  • Enzymology
  • Assay Development

Background:

  • Elastase is a key protease involved in various physiological and pathological processes.
  • Accurate and sensitive quantification of elastase activity is crucial for research and diagnostics.
  • Existing elastase assays may lack sensitivity, speed, or versatility.

Purpose of the Study:

  • To develop a novel, rapid, sensitive, and versatile assay for quantifying elastase activity.
  • To utilize succinylated elastin as a substrate for enhanced specificity and detection.
  • To enable high-throughput analysis of elastase in diverse biological matrices.

Main Methods:

  • Developed a colorimetric assay measuring primary amines exposed after protein degradation.
  • Employed succinylated elastin as the specific substrate for elastase.

Related Experiment Videos

  • Utilized trinitrobenzene sulfonic acid for quantifying the degree of protein digestion.
  • Performed assays in microtiter plate format for small sample volumes.

    • Main Results:

    • The assay is rapid and highly sensitive, detecting elastase down to 1 ng/ml.
    • Linearity of detection spans enzyme concentrations from 1 ng/ml to 10 microg/ml.
    • Succinylated elastin substrate demonstrated specificity for elastase over trypsin.
    • The assay is versatile, applicable to cell culture supernatants, plasma, biopsies, and homogenates.

    Conclusions:

    • A novel, rapid, sensitive, and versatile elastase assay has been successfully developed.
    • The assay offers high specificity for elastase using succinylated elastin and trinitrobenzene sulfonic acid.
    • Its microtiter plate format and applicability to various samples facilitate broad research use.