Specific enzymatic digestion and fragment isolation for the critical quality attribute assessment of IgG1

Rappo, Martin Alexander

doi:10.21256/zhaw-2059

Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-2059

Publication type:	Master thesis
Title:	Specific enzymatic digestion and fragment isolation for the critical quality attribute assessment of IgG1
Authors:	Rappo, Martin Alexander
Advisors / Reviewers:	Rohrer, Jack Rösli, Christoph
DOI:	10.21256/zhaw-2059
Extent:	57
Issue Date:	2017
Publisher / Ed. Institution:	ZHAW Zürcher Hochschule für Angewandte Wissenschaften
Language:	English
Subjects:	Critical Quality Attributes; CQA; Monoclonal Antibody; Mab; Fab; Fc; F(ab’)2; Large hinge lgG fragment; LHF; IdeS; Lysine Gingipain; Kgp; EndoS2; IgdE
Subject (DDC):	615: Pharmacology and therapeutics 660.6: Biotechnology
Abstract:	Recent ICH quality guideline updates introduced the enhanced drug development which is based on Quality by Design approach. For this a Quality Risk Management has to be conducted based on scientific knowledge. Information gaps are closed through analysis of molecules with domain-specific modifications. The aim of this thesis was to evaluate methods to prepare a model IgG1 for such studies. The antibody fragments F(ab’)2, LHF, Fab and Fc and the Fc-glycosylation cleaved IgG with residual N-acetylglucosamine were selected. A representative IgG1 was digested with the recently discovered enzymes IdeS, Kgp, IgdE and EndoS2 and the digestion products were purified with common biotechnological down-stream methods. The F(ab’)2 was successfully isolated with a yield of 55% and a purity of 96%. After an initial digestion parameter optimization the LHF, Fab and Fc could be generated and an isolation method was established. A following scale-up was unsuccessful, due to reproducibility issues. A promising alternative was identified. The enzymatic glycan cleavage and product isolation was successful with a yield of 80% and a purity of 99%.
URI:	https://digitalcollection.zhaw.ch/handle/11475/1871
License (according to publishing contract):	Licence according to publishing contract
Departement:	Life Sciences and Facility Management
Appears in collections:	Masterarbeiten Life Sciences

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Rappo, M. A. (2017). Specific enzymatic digestion and fragment isolation for the critical quality attribute assessment of IgG1 [Master’s thesis, ZHAW Zürcher Hochschule für Angewandte Wissenschaften]. https://doi.org/10.21256/zhaw-2059

Rappo, M.A. (2017) Specific enzymatic digestion and fragment isolation for the critical quality attribute assessment of IgG1. Master’s thesis. ZHAW Zürcher Hochschule für Angewandte Wissenschaften. Available at: https://doi.org/10.21256/zhaw-2059.

M. A. Rappo, “Specific enzymatic digestion and fragment isolation for the critical quality attribute assessment of IgG1,” Master’s thesis, ZHAW Zürcher Hochschule für Angewandte Wissenschaften, 2017. doi: 10.21256/zhaw-2059.

RAPPO, Martin Alexander, 2017. Specific enzymatic digestion and fragment isolation for the critical quality attribute assessment of IgG1. Master’s thesis. ZHAW Zürcher Hochschule für Angewandte Wissenschaften

Rappo, Martin Alexander. 2017. “Specific Enzymatic Digestion and Fragment Isolation for the Critical Quality Attribute Assessment of IgG1.” Master’s thesis, ZHAW Zürcher Hochschule für Angewandte Wissenschaften. https://doi.org/10.21256/zhaw-2059.

Rappo, Martin Alexander. Specific Enzymatic Digestion and Fragment Isolation for the Critical Quality Attribute Assessment of IgG1. ZHAW Zürcher Hochschule für Angewandte Wissenschaften, 2017, https://doi.org/10.21256/zhaw-2059.