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Characterization of the Chemical and Functional Stability of DEAE Sepharose Fast Flow
1993 (English)In: Process Biochemistry, ISSN 1359-5113, E-ISSN 1873-3298, Vol. 28, no 4, 223-230 p.Article in journal (Refereed) Published
Abstract [en]

The release of amines from the ion-exchange groups in DEAE Sepharose® Fast Flow has been studied under static and column conditions. The leakage compounds have been identified and quantified by gas chromatography—mass spectrometry. It was shown that the main leakage product under acidic (pH 1) and basic conditions (pH 14) was N,N,N′,N′-tetraethylethylenediamine. Three other amines were also identified, namely N,N,N′-triethylethylenediamine, diethylaminoethanol and diethylamine. The leakage of amines from DEAE Sepharose Fast Flow treated at pH 1 or 14 for 672 h at 40°C corresponds to a reduction of only 1% of the total ion-exchange capacity.

The functional stability of DEAE Sepharose Fast Flow was also studied by separation of a protein mixture during repeated cleaning-in-place treatments with 0·10 m HCl or 1·0 m NaOH. The separation behaviour was unaltered after the gel had been treated for a total contact time of 672 h with 0·10 m HCl or 1·0 m NaOH.

The clearance of ethanol from a DEAE Sepharose Fast Flow column stored in a 20% ethanol aqueous solution was also studied.

Place, publisher, year, edition, pages
1993. Vol. 28, no 4, 223-230 p.
National Category
Analytical Chemistry
URN: urn:nbn:se:uu:diva-234606DOI: 10.1016/0032-9592(93)80038-IOAI: oai:DiVA.org:uu-234606DiVA: diva2:757853
Available from: 2014-10-23 Created: 2014-10-21 Last updated: 2015-02-03Bibliographically approved
In thesis
1. Characterisation of Chromatography Media Aimed for Purification of Biomolecules
Open this publication in new window or tab >>Characterisation of Chromatography Media Aimed for Purification of Biomolecules
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Chromatography media (resins) are very important for and widely used by the biopharma industry in large scale production of biopharmaceuticals, e.g. monoclonal antibodies. Today there are several hundred biopharmaceuticals released globally on the healthcare market. This thesis discusses various strategies and methods for the characterisation of chemical and functional stability of chromatography media. In addition, various analytical techniques used in these areas were evaluated and applied. Further, more specific physical and chemical characterisation methods were evaluated and applied to explore different properties of various chromatography media.

In Papers I-III, established methodologies for performing chemical and functional stability studies were used. Mainly agarose-based chromatography media were investigated. For fast screening of the chemical stability, the total organic carbon analysis technique was evaluated and applied. This technique that measures the carbon leakage from the chromatography media at different conditions, proved to be very suitable and robust. For detection and/or identification of leakage compounds responsible for or for part of the measured carbon leakage, different methods such as (high performance) liquid chromatography and gas chromatography mass spectrometry were used.

In Papers IV-VII, different properties (i.e. functional performance, ligand content and surface chemistry) were evaluated for different agarose-based chromatography media. Standard chromatographic methods (ion exchange chromatography) and spectroscopic methods (e.g. Fourier transform infrared spectroscopy and time-of-flight secondary ion mass spectrometry) were evaluated and applied. Chemometric methods were used for efficient evaluation of data.

Information of chemical, functional and leakage data of chromatography media are valuable and important for the biopharmaceutical companies to be able to fulfil the regulatory requirements of biopharmaceuticals. In addition, information of various chemical, functional and physical properties of chromatography media is likewise important during development and set up of new biopharmaceutical processes.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2014. 73 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1202
Chromatography, Media, Characterisation, Stability
National Category
Analytical Chemistry
Research subject
Chemistry with specialization in Analytical Chemistry
urn:nbn:se:uu:diva-234743 (URN)978-91-554-9102-4 (ISBN)
Public defence
2014-12-19, B22, BMC, Uppsala, 13:15 (English)
Available from: 2014-11-28 Created: 2014-10-23 Last updated: 2015-02-03

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