Cargando…
Characterization of recombinant IgA producing CHO cell lines by qPCR
Immunoglobulin A (IgA) mediates a key role in mucosal immunity and is a promising novel immunotherapeutic candidate. However, difficulties in obtaining enough material often hamper in vivo explorations. We have previously generated recombinant Chinese hamster ovary (CHO) cell lines which expressed t...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3981580/ http://dx.doi.org/10.1186/1753-6561-7-S6-P114 |
_version_ | 1782311067725070336 |
---|---|
author | Reinhart, David Sommeregger, Wolfgang Debreczeny, Monika Gludovacz, Elisabeth Kunert, Renate |
author_facet | Reinhart, David Sommeregger, Wolfgang Debreczeny, Monika Gludovacz, Elisabeth Kunert, Renate |
author_sort | Reinhart, David |
collection | PubMed |
description | Immunoglobulin A (IgA) mediates a key role in mucosal immunity and is a promising novel immunotherapeutic candidate. However, difficulties in obtaining enough material often hamper in vivo explorations. We have previously generated recombinant Chinese hamster ovary (CHO) cell lines which expressed two different HIV-1 antibodies, 3D6 and 4B3, as IgA1 [1]. One cell line (3D6-IgA) shows high production rates, whereas the other (4B3-IgA) secretes rather low amounts of product. In order to unravel the mystery of productivity bottlenecks we extensively characterized the cell lines regarding growth rate, IgA productivity in long-term culture, immunofluorescence microscopy, flow cytometry and Western blotting of intra- and extracellular product (data not shown). The generated data encouraged us to analyze whether the observed antibody productivities could be explained by gene copy number (GCN) or mRNA levels. |
format | Online Article Text |
id | pubmed-3981580 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-39815802014-04-24 Characterization of recombinant IgA producing CHO cell lines by qPCR Reinhart, David Sommeregger, Wolfgang Debreczeny, Monika Gludovacz, Elisabeth Kunert, Renate BMC Proc Poster Presentation Immunoglobulin A (IgA) mediates a key role in mucosal immunity and is a promising novel immunotherapeutic candidate. However, difficulties in obtaining enough material often hamper in vivo explorations. We have previously generated recombinant Chinese hamster ovary (CHO) cell lines which expressed two different HIV-1 antibodies, 3D6 and 4B3, as IgA1 [1]. One cell line (3D6-IgA) shows high production rates, whereas the other (4B3-IgA) secretes rather low amounts of product. In order to unravel the mystery of productivity bottlenecks we extensively characterized the cell lines regarding growth rate, IgA productivity in long-term culture, immunofluorescence microscopy, flow cytometry and Western blotting of intra- and extracellular product (data not shown). The generated data encouraged us to analyze whether the observed antibody productivities could be explained by gene copy number (GCN) or mRNA levels. BioMed Central 2013-12-04 /pmc/articles/PMC3981580/ http://dx.doi.org/10.1186/1753-6561-7-S6-P114 Text en Copyright © 2013 Reinhart et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Poster Presentation Reinhart, David Sommeregger, Wolfgang Debreczeny, Monika Gludovacz, Elisabeth Kunert, Renate Characterization of recombinant IgA producing CHO cell lines by qPCR |
title | Characterization of recombinant IgA producing CHO cell lines by qPCR |
title_full | Characterization of recombinant IgA producing CHO cell lines by qPCR |
title_fullStr | Characterization of recombinant IgA producing CHO cell lines by qPCR |
title_full_unstemmed | Characterization of recombinant IgA producing CHO cell lines by qPCR |
title_short | Characterization of recombinant IgA producing CHO cell lines by qPCR |
title_sort | characterization of recombinant iga producing cho cell lines by qpcr |
topic | Poster Presentation |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3981580/ http://dx.doi.org/10.1186/1753-6561-7-S6-P114 |
work_keys_str_mv | AT reinhartdavid characterizationofrecombinantigaproducingchocelllinesbyqpcr AT sommereggerwolfgang characterizationofrecombinantigaproducingchocelllinesbyqpcr AT debreczenymonika characterizationofrecombinantigaproducingchocelllinesbyqpcr AT gludovaczelisabeth characterizationofrecombinantigaproducingchocelllinesbyqpcr AT kunertrenate characterizationofrecombinantigaproducingchocelllinesbyqpcr |