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Surface Chemical Analysis of Carbohydrate Materials Used for Chromatography Media by Time-of-Flight Secondary Ion Mass Spectrometry
2004 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 76, no 7, 1857-1864 p.Article in journal (Refereed) Published
Abstract [en]

The surface chemical structure of two raw materials (agarose and dextran) and four base matrixes used in the manufacture of chromatography media were analyzed using time-of-flight secondary ion mass spectrometry (TOF-SIMS). The results show that the small differences in molecular structure between these materials result in significant differences in the TOF-SIMS spectra and that these differences can be identified and quantified using either of two different approaches. In a novel approach, fragment ion distributions were extracted from the TOF-SIMS spectra for each material, providing an immediate and systematic overview of the spectral features. Difference fragment distributions were used to highlight spectral differences between the materials. The results of the fragment ion distribution analysis, in terms of identification and quantification of spectral variations between different materials, were found to be in agreement with the results from a principal component analysis using the same set of data. Both methods were found capable of (i) distinguishing between agarose and dextran and (ii) detecting and quantifying the degree of cross-linking present in the four base matrix materials. In addition, using a deuterated chemical cross-linker, it was possible to identify spectral features specifically connected to the cross-link molecular structure.

Place, publisher, year, edition, pages
2004. Vol. 76, no 7, 1857-1864 p.
National Category
Analytical Chemistry
URN: urn:nbn:se:uu:diva-234612DOI: 10.1021/ac035457gPubMedID: 15053644OAI: oai:DiVA.org:uu-234612DiVA: diva2:757860
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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