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Related Experiment Video

Updated: Apr 13, 2026

High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes
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[Characterization and comparison of interferon reference standards using UPLC-MS].

Lei Tao, De-ning Pei, Chun-mei Han

    Yao Xue Xue Bao = Acta Pharmaceutica Sinica
    |May 1, 2015
    PubMed
    Summary
    This summary is machine-generated.

    This study compared interferon reference standards from five manufacturers. Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) confirmed amino acid sequences and identified critical post-translational modifications, ensuring quality control for these vital biopharmaceuticals.

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    Area of Science:

    • Biopharmaceutical Analysis
    • Protein Chemistry
    • Analytical Chemistry

    Background:

    • Interferons are crucial biopharmaceuticals with complex structures.
    • Ensuring the quality and consistency of interferon reference standards is vital for accurate therapeutic drug monitoring and research.
    • Variations in post-translational modifications can impact interferon efficacy and safety.

    Purpose of the Study:

    • To characterize and compare interferon reference standards from multiple manufacturers.
    • To verify the amino acid sequence and identify post-translational modifications of interferon standards.
    • To evaluate the suitability of Ultra-performance Liquid Chromatography-Mass Spectrometry (UPLC-MS) for interferon standard characterization.

    Main Methods:

    • Analysis of molecular mass using mass spectrometry.
    • Trypsin-digested peptide mass mapping for amino acid sequence verification.
    • Identification of post-translational modifications, including disulfide bonds, N-terminal modifications, and methionine oxidation using UPLC-MS.

    Main Results:

    • Molecular mass and amino acid sequences of interferon standards were consistent with theoretical values.
    • Disulfide bond patterns varied between lots: Cys1-Cys98/Cys29-Cys138 in four lots and Cys29-Cys139/Cys86-Cys99 in one lot.
    • Post-translational modifications such as N-terminal "+Met", N-terminal acetylation, and methionine oxidation were detected in a subset of samples.

    Conclusions:

    • UPLC-MS is a powerful technique for comprehensive characterization of interferon reference standards.
    • The study identified variations in disulfide bonds and other post-translational modifications among different interferon lots.
    • These findings highlight the importance of rigorous characterization for ensuring the quality and comparability of interferon reference standards across manufacturers.