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Related Experiment Videos

Competitive protein-binding assay for trimetrexate.

J C Drake, C J Allegra, G A Curt

    Cancer Treatment Reports
    |June 1, 1985
    PubMed
    Summary
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    A new assay accurately measures trimetrexate (TMTX) levels in biological fluids. This method utilizes trimetrexate

    Area of Science:

    • Biochemistry
    • Pharmacology
    • Analytical Chemistry

    Background:

    • Trimetrexate (TMTX) is an antifolate drug that inhibits dihydrofolate reductase.
    • Accurate quantification of TMTX in biological samples is crucial for therapeutic drug monitoring.

    Purpose of the Study:

    • To develop and validate a competitive protein-binding assay for trimetrexate (TMTX).
    • To enable precise measurement of TMTX concentrations in various biological matrices.

    Main Methods:

    • A competitive protein-binding assay was designed using human dihydrofolate reductase (hDHFR).
    • Trimetrexate (TMTX) competed with radiolabeled methotrexate (3H-methotrexate) for binding to hDHFR.
    • Dextran-albumin-coated charcoal was used to separate bound drug from free drug.

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    Main Results:

    • The assay demonstrated sensitivity for TMTX in the range of 2 x 10(-9) to 5 x 10(-8) M.
    • Intra-assay variability was low, with a coefficient of variation below 10%.
    • Measurements in plasma, cerebrospinal fluid, and urine showed close agreement with aqueous standard determinations.

    Conclusions:

    • A reliable competitive protein-binding assay for trimetrexate (TMTX) has been established.
    • This assay is suitable for quantifying TMTX in clinical samples like plasma, CSF, and urine.
    • The developed assay facilitates therapeutic drug monitoring of trimetrexate.