| Parameters to enhance human hybridoma formation with hypoosmolar electrofusion. | |
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MedLine Citation:
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PMID: 1651789 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Hypoosmolar conditions have permitted the development of electrofusion techniques capable of producing human hybridomas from as few as 10(5) B cells. A hybridoma formation efficiency of one hybrid for each 125 input B cells has been achieved with Epstein-Barr virus--activated B cells and mouse-human heteromyelomas. This is at least 100-fold higher in efficiency than with polyethylene glycol-induced cell fusion, as well as a 50- to 100-fold decrease in the required number of human B cells. The ability to fuse a small number of input B cells should lead to a greater success rate in immortalizing the rare antigen-specific B cells. The critical parameters include fusion voltages, the composition and number of wash steps used in cell preparation, the composition and duration of exposure to hypoosmolar fusion medium, fusion ratio, plating density, the use of growth medium without pH indicator, and the use of an irradiated human fibroblast feeder layer. By manipulating these parameters, a high hybrid yield can be achieved with different mouse-human heteromyelomas and Epstein-Barr virus-activated B cells. |
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Authors:
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S Perkins; U Zimmermann; S K Foung |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Human antibodies and hybridomas Volume: 2 ISSN: 0956-960X ISO Abbreviation: Hum. Antibodies Hybridomas Publication Date: 1991 Jul |
Date Detail:
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Created Date: 1991-09-24 Completed Date: 1991-09-24 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 9014461 Medline TA: Hum Antibodies Hybridomas Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 155-9 Citation Subset: IM |
Affiliation:
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Department of Pathology, Stanford University School of Medicine, CA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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B-Lymphocytes
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immunology Culture Media Herpesvirus 4, Human Humans Hybridomas* Lymphocyte Activation Methods Osmolar Concentration |
| Grant Support | |
ID/Acronym/Agency:
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AI22557/AI/NIAID NIH HHS; AI26031/AI/NIAID NIH HHS; HL33811/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Culture Media |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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