کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2085016 1545413 2006 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Determination of Pulmozyme® (dornase alpha) stability using a kinetic colorimetric DNase I activity assay
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Determination of Pulmozyme® (dornase alpha) stability using a kinetic colorimetric DNase I activity assay
چکیده انگلیسی

An enzymatic activity assay was developed for the determination of dornase alpha human recombinant desoxyribonuclease (DNase I) stability. The method was adapted from a colorimetric endpoint enzyme activity assay for DNase I based on the degradation of a DNA/methyl green complex. With the described modifications the kinetic measurement of enzyme activity is feasible on an automated analyzer system within a rather short time. The development of this assay was based on the need for reliable detection of a possible loss of enzyme activity after transferring the commercial therapeutic agent into sealed glass vials required for a placebo-controlled study. The measuring range of this stability test was from 0 to 3000 U/L corresponding to 0–120% of the original enzyme activity; CV values of control solutions inside the measuring range were between 3% and 5%. The enzyme activity decreased less than 15% during the observation period of 180 days. In conclusion the current kinetic assay is a reliable method for a simple time-saving determination of DNase I activity to test Pulmozyme® stability as required for quality control. As dornase alpha is used for inhalation, this method also proved its reliability in testing DNase stability during aerosolization with new inhalation devices (e-flow).

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: European Journal of Pharmaceutics and Biopharmaceutics - Volume 63, Issue 3, July 2006, Pages 365–368
نویسندگان
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