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A novel method for making human monoclonal antibodies
We have developed a B cell immortalization method for low B cell numbers per well using simultaneous B cell stimulation by CpG2006 and B cell infection by Epstein-Barr virus (EBV), followed by an additional CpG2006 and interleukin-2 (IL-2) stimulus. Using this method, immunoglobulin G (IgG)-producin...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier Ltd.
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7126289/ https://www.ncbi.nlm.nih.gov/pubmed/20732843 http://dx.doi.org/10.1016/j.jaut.2010.05.001 |
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author | Fraussen, J. Vrolix, K. Martinez-Martinez, P. Losen, M. Meulemans, E. De Baets, M.H. Stinissen, P. Somers, V. |
author_facet | Fraussen, J. Vrolix, K. Martinez-Martinez, P. Losen, M. Meulemans, E. De Baets, M.H. Stinissen, P. Somers, V. |
author_sort | Fraussen, J. |
collection | PubMed |
description | We have developed a B cell immortalization method for low B cell numbers per well using simultaneous B cell stimulation by CpG2006 and B cell infection by Epstein-Barr virus (EBV), followed by an additional CpG2006 and interleukin-2 (IL-2) stimulus. Using this method, immunoglobulin G (IgG)-producing immortalized B cell lines were generated from peripheral blood IgG(+)CD22(+) B cells with an efficiency of up to 83%. Antibody can already be obtained from the culture supernatant after 3–4 weeks. Moreover, clonality analysis demonstrated monoclonality in 87% of the resulting immortalized B cell lines. Given the high immortalization efficiency and monoclonality rate, evidence is provided that no further subcloning is necessary. An important application of this B cell immortalization method is the characterization of (autoreactive) antibodies from patients with autoimmune disease. This could eventually lead to the identification of new autoantigens, disease markers or targets for therapy. |
format | Online Article Text |
id | pubmed-7126289 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Elsevier Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71262892020-04-08 A novel method for making human monoclonal antibodies Fraussen, J. Vrolix, K. Martinez-Martinez, P. Losen, M. Meulemans, E. De Baets, M.H. Stinissen, P. Somers, V. J Autoimmun Article We have developed a B cell immortalization method for low B cell numbers per well using simultaneous B cell stimulation by CpG2006 and B cell infection by Epstein-Barr virus (EBV), followed by an additional CpG2006 and interleukin-2 (IL-2) stimulus. Using this method, immunoglobulin G (IgG)-producing immortalized B cell lines were generated from peripheral blood IgG(+)CD22(+) B cells with an efficiency of up to 83%. Antibody can already be obtained from the culture supernatant after 3–4 weeks. Moreover, clonality analysis demonstrated monoclonality in 87% of the resulting immortalized B cell lines. Given the high immortalization efficiency and monoclonality rate, evidence is provided that no further subcloning is necessary. An important application of this B cell immortalization method is the characterization of (autoreactive) antibodies from patients with autoimmune disease. This could eventually lead to the identification of new autoantigens, disease markers or targets for therapy. Elsevier Ltd. 2010-09 2010-06-08 /pmc/articles/PMC7126289/ /pubmed/20732843 http://dx.doi.org/10.1016/j.jaut.2010.05.001 Text en Copyright © 2010 Elsevier Ltd. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Fraussen, J. Vrolix, K. Martinez-Martinez, P. Losen, M. Meulemans, E. De Baets, M.H. Stinissen, P. Somers, V. A novel method for making human monoclonal antibodies |
title | A novel method for making human monoclonal antibodies |
title_full | A novel method for making human monoclonal antibodies |
title_fullStr | A novel method for making human monoclonal antibodies |
title_full_unstemmed | A novel method for making human monoclonal antibodies |
title_short | A novel method for making human monoclonal antibodies |
title_sort | novel method for making human monoclonal antibodies |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7126289/ https://www.ncbi.nlm.nih.gov/pubmed/20732843 http://dx.doi.org/10.1016/j.jaut.2010.05.001 |
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