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Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter

Most therapeutic monoclonal antibodies in biopharmaceutical processes are produced in Chinese hamster ovary (CHO) cells. Technological advances have rendered the selection procedure for higher producers a robust protocol. However, information on molecular mechanisms that impart the property of hyper...

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Autores principales: Dahodwala, Hussain, Kaushik, Prashant, Tejwani, Vijay, Kuo, Chih-Chung, Menard, Patrice, Henry, Michael, Voldborg, Bjorn G., Lewis, Nathan E., Meleady, Paula, Sharfstein, Susan T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7311070/
https://www.ncbi.nlm.nih.gov/pubmed/32577618
http://dx.doi.org/10.1016/j.crbiot.2019.09.001
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author Dahodwala, Hussain
Kaushik, Prashant
Tejwani, Vijay
Kuo, Chih-Chung
Menard, Patrice
Henry, Michael
Voldborg, Bjorn G.
Lewis, Nathan E.
Meleady, Paula
Sharfstein, Susan T.
author_facet Dahodwala, Hussain
Kaushik, Prashant
Tejwani, Vijay
Kuo, Chih-Chung
Menard, Patrice
Henry, Michael
Voldborg, Bjorn G.
Lewis, Nathan E.
Meleady, Paula
Sharfstein, Susan T.
author_sort Dahodwala, Hussain
collection PubMed
description Most therapeutic monoclonal antibodies in biopharmaceutical processes are produced in Chinese hamster ovary (CHO) cells. Technological advances have rendered the selection procedure for higher producers a robust protocol. However, information on molecular mechanisms that impart the property of hyper-productivity in the final selected clones is currently lacking. In this study, an IgG-producing industrial cell line and its methotrexate (MTX)-amplified progeny cell line were analyzed using transcriptomic, proteomic, phosphoproteomic, and chromatin immunoprecipitation (ChIP) techniques. Computational prediction of transcription factor binding to the transgene cytomegalovirus (CMV) promoter by the Transcription Element Search System and upstream regulator analysis of the differential transcriptomic data suggested increased in vivo CMV promoter-cAMP response element binding protein (CREB1) interaction in the higher producing cell line. Differential nuclear proteomic analysis detected 1.3-fold less CREB1 in the nucleus of the high productivity cell line compared with the parental cell line. However, the differential abundance of multiple CREB1 phosphopeptides suggested an increase in CREB1 activity in the higher producing cell line, which was confirmed by increased association of the CMV promotor with CREB1 in the high producer cell line. Thus, we show here that the nuclear proteome and phosphoproteome have an important role in regulating final productivity of recombinant proteins from CHO cells, and that CREB1 may play a role in transcriptional enhancement. Moreover, CREB1 phosphosites may be potential targets for cell engineering for increased productivity.
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spelling pubmed-73110702020-06-23 Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter Dahodwala, Hussain Kaushik, Prashant Tejwani, Vijay Kuo, Chih-Chung Menard, Patrice Henry, Michael Voldborg, Bjorn G. Lewis, Nathan E. Meleady, Paula Sharfstein, Susan T. Curr Res Biotechnol Article Most therapeutic monoclonal antibodies in biopharmaceutical processes are produced in Chinese hamster ovary (CHO) cells. Technological advances have rendered the selection procedure for higher producers a robust protocol. However, information on molecular mechanisms that impart the property of hyper-productivity in the final selected clones is currently lacking. In this study, an IgG-producing industrial cell line and its methotrexate (MTX)-amplified progeny cell line were analyzed using transcriptomic, proteomic, phosphoproteomic, and chromatin immunoprecipitation (ChIP) techniques. Computational prediction of transcription factor binding to the transgene cytomegalovirus (CMV) promoter by the Transcription Element Search System and upstream regulator analysis of the differential transcriptomic data suggested increased in vivo CMV promoter-cAMP response element binding protein (CREB1) interaction in the higher producing cell line. Differential nuclear proteomic analysis detected 1.3-fold less CREB1 in the nucleus of the high productivity cell line compared with the parental cell line. However, the differential abundance of multiple CREB1 phosphopeptides suggested an increase in CREB1 activity in the higher producing cell line, which was confirmed by increased association of the CMV promotor with CREB1 in the high producer cell line. Thus, we show here that the nuclear proteome and phosphoproteome have an important role in regulating final productivity of recombinant proteins from CHO cells, and that CREB1 may play a role in transcriptional enhancement. Moreover, CREB1 phosphosites may be potential targets for cell engineering for increased productivity. 2019-10-05 2019-11 /pmc/articles/PMC7311070/ /pubmed/32577618 http://dx.doi.org/10.1016/j.crbiot.2019.09.001 Text en Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
spellingShingle Article
Dahodwala, Hussain
Kaushik, Prashant
Tejwani, Vijay
Kuo, Chih-Chung
Menard, Patrice
Henry, Michael
Voldborg, Bjorn G.
Lewis, Nathan E.
Meleady, Paula
Sharfstein, Susan T.
Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter
title Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter
title_full Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter
title_fullStr Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter
title_full_unstemmed Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter
title_short Increased mAb production in amplified CHO cell lines is associated with increased interaction of CREB1 with transgene promoter
title_sort increased mab production in amplified cho cell lines is associated with increased interaction of creb1 with transgene promoter
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7311070/
https://www.ncbi.nlm.nih.gov/pubmed/32577618
http://dx.doi.org/10.1016/j.crbiot.2019.09.001
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