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Germinality does not necessarily define mAb expression and thermal stability

The production potential of recombinant monoclonal antibody (mAb) expressing cell lines depends, among other factors, on the intrinsic antibody structure determined by the amino acid sequence. In this study, we investigated the influence of somatic mutations in the V(D)J sequence of four individual,...

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Autores principales: Schwaigerlehner, Linda, Mayrhofer, Patrick, Diem, Matthias, Steinfellner, Willibald, Fenech, Emma, Oostenbrink, Chris, Kunert, Renate
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719414/
https://www.ncbi.nlm.nih.gov/pubmed/31350616
http://dx.doi.org/10.1007/s00253-019-09998-3
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author Schwaigerlehner, Linda
Mayrhofer, Patrick
Diem, Matthias
Steinfellner, Willibald
Fenech, Emma
Oostenbrink, Chris
Kunert, Renate
author_facet Schwaigerlehner, Linda
Mayrhofer, Patrick
Diem, Matthias
Steinfellner, Willibald
Fenech, Emma
Oostenbrink, Chris
Kunert, Renate
author_sort Schwaigerlehner, Linda
collection PubMed
description The production potential of recombinant monoclonal antibody (mAb) expressing cell lines depends, among other factors, on the intrinsic antibody structure determined by the amino acid sequence. In this study, we investigated the influence of somatic mutations in the V(D)J sequence of four individual, mature model mAbs on the expression potential. Therefore, we defined four couples, each consisting of one naturally occurring mAb (2G12, Ustekinumab, 4B3, and 2F5) and the corresponding germline-derived cognate mAb (353/11, 554/12, 136/63, and 236/14). For all eight mAb variants, recombinant Chinese hamster ovary (CHO) cell lines were developed with mAbs expressed from a defined chromosomal locus. The presented workflow investigates critical parameters including productivity, intra- and extracellular product profile, XBP1 splicing, thermal stability, and in silico hydrophobicity. Significant differences in productivity were even observed between the germline-derived mAbs which did not undergo somatic mutagenesis. Accordingly, back-to-germline mutations of mature mAbs are not necessarily reflecting improved expression and stability but indicate opportunities and limits of mAb engineering. From our studies, we conclude that germinalization represents a potential to improve mAb properties depending on the antibody’s germline family, highlighting the fact that mAbs should be treated individually. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-019-09998-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-67194142019-09-19 Germinality does not necessarily define mAb expression and thermal stability Schwaigerlehner, Linda Mayrhofer, Patrick Diem, Matthias Steinfellner, Willibald Fenech, Emma Oostenbrink, Chris Kunert, Renate Appl Microbiol Biotechnol Biotechnological Products and Process Engineering The production potential of recombinant monoclonal antibody (mAb) expressing cell lines depends, among other factors, on the intrinsic antibody structure determined by the amino acid sequence. In this study, we investigated the influence of somatic mutations in the V(D)J sequence of four individual, mature model mAbs on the expression potential. Therefore, we defined four couples, each consisting of one naturally occurring mAb (2G12, Ustekinumab, 4B3, and 2F5) and the corresponding germline-derived cognate mAb (353/11, 554/12, 136/63, and 236/14). For all eight mAb variants, recombinant Chinese hamster ovary (CHO) cell lines were developed with mAbs expressed from a defined chromosomal locus. The presented workflow investigates critical parameters including productivity, intra- and extracellular product profile, XBP1 splicing, thermal stability, and in silico hydrophobicity. Significant differences in productivity were even observed between the germline-derived mAbs which did not undergo somatic mutagenesis. Accordingly, back-to-germline mutations of mature mAbs are not necessarily reflecting improved expression and stability but indicate opportunities and limits of mAb engineering. From our studies, we conclude that germinalization represents a potential to improve mAb properties depending on the antibody’s germline family, highlighting the fact that mAbs should be treated individually. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-019-09998-3) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2019-07-26 2019 /pmc/articles/PMC6719414/ /pubmed/31350616 http://dx.doi.org/10.1007/s00253-019-09998-3 Text en © The Author(s) 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Biotechnological Products and Process Engineering
Schwaigerlehner, Linda
Mayrhofer, Patrick
Diem, Matthias
Steinfellner, Willibald
Fenech, Emma
Oostenbrink, Chris
Kunert, Renate
Germinality does not necessarily define mAb expression and thermal stability
title Germinality does not necessarily define mAb expression and thermal stability
title_full Germinality does not necessarily define mAb expression and thermal stability
title_fullStr Germinality does not necessarily define mAb expression and thermal stability
title_full_unstemmed Germinality does not necessarily define mAb expression and thermal stability
title_short Germinality does not necessarily define mAb expression and thermal stability
title_sort germinality does not necessarily define mab expression and thermal stability
topic Biotechnological Products and Process Engineering
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719414/
https://www.ncbi.nlm.nih.gov/pubmed/31350616
http://dx.doi.org/10.1007/s00253-019-09998-3
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