کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1198903 1493489 2016 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Application of spectral deconvolution and inverse mechanistic modelling as a tool for root cause investigation in protein chromatography
ترجمه فارسی عنوان
کاربرد تقسیم بندی طیفی و مدلسازی مکانیکی معکوس به عنوان یک ابزار برای بررسی ریشه در کروماتوگرافی پروتئین
کلمات کلیدی
تحقیق علت ریشه، تکنولوژی تحلیلی فرآیند، نظارت داخلی، مدل سازی مکانیکی، رگرسیون حداقل مربعات جزئی، مقدار کمی پروتئین انتخابی
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• A tool for rapid root cause investigation in protein chromatography was developed.
• The tool is based on a combination of spectral deconvolution and inverse mechanistic chromatography modelling.
• The tool is capable of identifying the causes for deviations in a chromatogram.
• This allows for a correction of a detected problem in the next cycle or batch.

In chromatographic protein purification, process variations, aging of columns, or processing errors can lead to deviations of the expected elution behavior of product and contaminants and can result in a decreased pool purity or yield. A different elution behavior of all or several involved species leads to a deviating chromatogram. The causes for deviations are however hard to identify by visual inspection and complicate the correction of a problem in the next cycle or batch. To overcome this issue, a tool for root cause investigation in protein chromatography was developed. The tool combines a spectral deconvolution with inverse mechanistic modelling. Mid-UV spectral data and Partial Least Squares Regression were first applied to deconvolute peaks to obtain the individual elution profiles of co-eluting proteins. The individual elution profiles were subsequently used to identify errors in process parameters by curve fitting to a mechanistic chromatography model. The functionality of the tool for root cause investigation was successfully demonstrated in a model protein study with lysozyme, cytochrome c, and ribonuclease A. Deviating chromatograms were generated by deliberately caused errors in the process parameters flow rate and sodium-ion concentration in loading and elution buffer according to a design of experiments. The actual values of the three process parameters and, thus, the causes of the deviations were estimated with errors of less than 4.4%. Consequently, the established tool for root cause investigation is a valuable approach to rapidly identify process variations, aging of columns, or processing errors. This might help to minimize batch rejections or contribute to an increased productivity.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1437, 11 March 2016, Pages 158–167
نویسندگان
, , ,