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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,...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2019
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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. |
format | Online Article Text |
id | pubmed-6719414 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
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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