Linked Life Data

12709022

Source:http://linkedlifedata.com/resource/pubmed/id/12709022

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pubmed-article:12709022pubmed:abstractTextWhereas functional heavy (H)-chain antibodies devoid of light (L)- chains account for about half of the circulating immunoglobulins in Camelidae, H-chain only antibodies (HCAbs) are not produced in other healthy mammals including rodents and humans. To test the feasibility of expressing single chain antibodies in the mouse, which on account of their small size and antigen-recognition properties would have a major impact on antibody engineering strategies, we constructed a rearranged dromedary H-chain gene encoding the immunoglobulin G2a (IgG2a) isotype with specificity for hen-egg lysozyme (HEL). This IgG2a H-chain gene was introduced into mouse myeloma cells not expressing endogenous immunoglobulin H- or L-chains. Unexpectedly the mouse cells processed and expressed the introduced H-chain as naturally occurring dromedary antibody. For this the first constant (C) region exon was proficiently removed from the recombinant H-chain transcript. This resulted in specific H-chain antibodies of the correct molecular weight (2 x 50 000 MW) secreted as disulfide-linked homodimers and displayed on the mouse cell surface as glycosyl-phosphatidyl-inositol-linked B-cell receptor. The results indicate that antibody expression and maturation without immunoglobulin L-chain is feasible and paves the way for the generation of transgenic single chain antibody repertoires.lld:pubmed
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pubmed-article:12709022pubmed:authorpubmed-author:BrysLeaLlld:pubmed
pubmed-article:12709022pubmed:authorpubmed-author:ZouXiangangXlld:pubmed
pubmed-article:12709022pubmed:authorpubmed-author:MuyldermansSe...lld:pubmed
pubmed-article:12709022pubmed:authorpubmed-author:NguyenViet...lld:pubmed
pubmed-article:12709022pubmed:authorpubmed-author:LauwereysMarc...lld:pubmed
pubmed-article:12709022pubmed:authorpubmed-author:BrüggemannMar...lld:pubmed
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pubmed-article:12709022pubmed:dateRevised2009-11-18lld:pubmed
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pubmed-article:12709022pubmed:year2003lld:pubmed
pubmed-article:12709022pubmed:articleTitleHeavy-chain only antibodies derived from dromedary are secreted and displayed by mouse B cells.lld:pubmed
pubmed-article:12709022pubmed:affiliationDepartment of Ultrastructure, Vlaams Interuniversitair Instituut voor Biotechnologie, Vrije Universiteit Brussel, Belgium.lld:pubmed
pubmed-article:12709022pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:12709022pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed