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Characterization of monoclonal antibodies (mAb) using high resolving analytical techniques
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
2019 (English)Independent thesis Advanced level (professional degree), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

In monoclonal antibody (mAb) production, multiple variants are introduced. These changes will affect the final product and due to product heterogeneity it is of importance to monitor these to ensure final product quality. In this master thesis, a multi attribute monitoring method by mass spectrometry (MAM-MS) was developed and then applied on a large bioreactor sample set. The goal with this project was to develop, evaluate and apply the developed method on mAb bioreactor samples. A purification method using PreDictor MabSelectTM PrismA affinity capture followed by a digestion method was developed and optimized. Both the affinity capture and tryptic digestion methods showed to be robust and reproducible. Two LC-MS/MS methods were developed, one data dependent acquisition method for identification of post-translational modifications (PTM) and one data independent acquisition method for quantification of PTMs. Using the developed MAM-MS method, identification and quantification of modifications of interest was possible. The mAb standards used during the whole all experiments during this project indicates that the approach is robust and reproducible. The technique showed promising results, indicating that there is a possibility for future implementation of the techniqe at GEHC. Further work on the method and a deeper comparison to currently used methods are required, to ensure that the MAM-MS method can replace the classical methods that are used today for PQA analysis of mAbs.

Place, publisher, year, edition, pages
2019. , p. 48
Series
UPTEC X ; 19 033
National Category
Engineering and Technology Biological Sciences
Identifiers
URN: urn:nbn:se:uu:diva-388536OAI: oai:DiVA.org:uu-388536DiVA, id: diva2:1333685
Educational program
Molecular Biotechnology Engineering Programme
Presentation
2019-06-04, BMC, Uppsala, 11:00 (English)
Supervisors
Examiners
Available from: 2019-07-02 Created: 2019-07-01 Last updated: 2019-07-02Bibliographically approved

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