Journal article
Generation of a human monoclonal antibody to hepatitis C virus, JRA1 by activation of peripheral blood lymphocytes and hypo-osmolar electrofusion
Human antibodies, Vol.6(2), pp.77-80
06/01/1995
DOI: 10.3233/HAB-1995-6207
PMID: 7492755
Abstract
We have generated a human monoclonal antibody with binding specificity for hepatitis C virus (HCV)-specific peptides using peripheral blood lymphocytes isolated from a HCV antibody positive patient. The B-lymphocytes were stimulated with lipopolysaccharide (LPS) for 72 hours prior to the fusion. A recently described high efficiency hypo-osmolar electrofusion technique was employed, allowing generation of a large number of human hybridomas. The hybridomas were screened for human immunoglobulin and HCV-specific peptide binding by EIA. A single HCV-positive clone, JRA1, was detected and sub-cloned. Isotype analysis showed it to secrete an IgM lambda monoclonal antibody. The antibody was positive on both first and second generation HCV antibody analysis. This study confirms that viable pathogen-specific B-cells may be recovered from the peripheral blood. Although such cells are likely to be relatively uncommon in the circulating B-cell pool, they may be successfully immortalized by high efficiency electrofusion techniques. This technique might be valuable for the generation of human monoclonal antibodies with specificity for other human pathogens.
Details
- Title: Subtitle
- Generation of a human monoclonal antibody to hepatitis C virus, JRA1 by activation of peripheral blood lymphocytes and hypo-osmolar electrofusion
- Creators
- Ulrich Zimmermann - Lehrstuhl für Biotechnologie, Biozentrum, Universität Würzburg, Würzburg, Germany D-97074Laurie Love-Homan - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA 52242Petra Gessner - Lehrstuhl für Biotechnologie, Biozentrum, Universität Würzburg, Würzburg, Germany D-97074Deborah Clark - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA 52242Gerd Klöck - Lehrstuhl für Biotechnologie, Biozentrum, Universität Würzburg, Würzburg, Germany D-97074Frederick C Johlin - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA 52242Garry A Neil - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA 52242
- Resource Type
- Journal article
- Publication Details
- Human antibodies, Vol.6(2), pp.77-80
- DOI
- 10.3233/HAB-1995-6207
- PMID
- 7492755
- ISSN
- 1093-2607
- eISSN
- 1875-869X
- Language
- English
- Date published
- 06/01/1995
- Academic Unit
- Gastroenterology and Hepatology; Internal Medicine
- Record Identifier
- 9984094873602771
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