Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-2059
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
Publisher / Ed. Institution : ZHAW Zürcher Hochschule für Angewandte Wissenschaften
Issue Date: 2017
Language : Englisch / 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: Pharmakologie und Therapeutik
660.6: Biotechnologie
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%.
Departement: Life Sciences und Facility Management
Publication type: Thesis: Master / Master Thesis
DOI : 10.21256/zhaw-2059
URI: https://digitalcollection.zhaw.ch/handle/11475/1871
Appears in Collections:Masterarbeiten Life Sciences

Files in This Item:
File Description SizeFormat 
2017_Rappo_Martin_MA_MScLS_PB.pdf7.25 MBAdobe PDFThumbnail
View/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.