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Improved lysozyme assay in biological fluids.

N Grossowicz, M Ariel, T Weber

    Clinical Chemistry
    |March 1, 1979
    PubMed
    Summary
    This summary is machine-generated.

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    This study presents a simple, rapid, and reproducible assay for lysozyme using Micrococcus lysodeikticus cell suspensions. The assay accurately quantifies lysozyme activity by measuring absorbance changes, detecting concentrations as low as 0.02 mg/L.

    Area of Science:

    • Biochemistry
    • Enzymology
    • Assay Development

    Background:

    • Lysozyme is a crucial enzyme in innate immunity and bacterial cell wall degradation.
    • Accurate and sensitive assays are essential for studying lysozyme's biological roles and therapeutic applications.
    • Existing lysozyme assays may lack simplicity, reproducibility, or sensitivity.

    Purpose of the Study:

    • To develop and validate a simple, rapid, sensitive, and highly reproducible assay for quantifying lysozyme activity.
    • To establish a stable substrate for long-term use in lysozyme assays.
    • To determine the lower limit of detection for egg-white lysozyme using the developed assay.

    Main Methods:

    • Utilized concentrated cell suspensions of Micrococcus lysodeikticus in a Tris-buffered glycerol/water solution (pH 7.5) as the substrate.

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  • Stored substrate cells at -20°C to ensure long-term stability of their susceptibility to lysozyme.
  • Measured lysozyme activity by monitoring the decrease in absorbance at 37°C after a 15-minute incubation period.

    • Main Results:

    • The developed assay demonstrated high reproducibility, with consistent concentration curves over an eight-month period.
    • The assay is sensitive, capable of accurately quantifying egg-white lysozyme concentrations as low as 0.02 mg/L.
    • Substrate cell stability was maintained for extended periods when stored at -20°C.

    Conclusions:

    • A simple, rapid, sensitive, and reproducible lysozyme assay has been successfully developed.
    • The use of Tris-buffered glycerol/water-suspended Micrococcus lysodeikticus cells provides a stable and reliable substrate for lysozyme quantification.
    • This assay is suitable for accurately measuring low concentrations of lysozyme, facilitating research in various biological contexts.