A retrovirus-based protein complementation assay screen reveals functional AKT1-binding partners

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A retrovirus-based protein complementation assay screen reveals functional AKT1-binding partners

Publisher
National Academy of Sciences
Publication Date
Oct 03, 2006
Source
PMC
Keywords
Disciplines
  • Medicine
License
Unknown

Abstract

Generation of heavy-chain-only antibodies in mice Rick Janssens*, Sylvia Dekker*, Rudi W. Hendriks†, George Panayotou‡, Alexandra van Remoortere§, John Kong-a San*, Frank Grosveld*¶, and Dubravka Drabek* *Departments of Cell Biology and †Immunology, ErasmusMC, P.O. Box 2040, 3000 CA, Rotterdam, The Netherlands; ‡Biomedical Sciences Research Center, Alexander Fleming, Varkiza 16602, Greece; and §Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands Edited by Richard A. Flavell, Yale University School of Medicine, New Haven, CT, and approved August 15, 2006 (received for review February 9, 2006) We have generated transgenic mice containing hybrid llama� human antibody loci that contain two llama variable regions and the human D, J, and C� and�or C� constant regions. Such loci rearrange productively and rescue B cell development efficiently without LC rearrangement. Heavy-chain-only antibodies (HCAb) are expressed at high levels, provided that the CH1 domain is deleted from the constant regions. HCAb production does not require an IgM stage for effective pre-B cell signaling. Antigen- specific heavy-chain-only IgM or IgGs are produced upon immuni- zation. The IgG is dimeric, whereas IgM is multimeric. The chimeric HCAb loci are subject to allelic exclusion, but several copies of the transgenic locus can be rearranged and expressed successfully on the same allele in the same cell. Such cells are not subject to negative selection. The mice produce a full antibody repertoire and provide a previously undescribed avenue to produce specific hu- man HCAb in the future. immunoglobulin rearrangement � transgenic Conventional antibodies contain two heavy and light chains (LC)coded for by heavy and LC loci. B cell development and antibody production starts in the bonemarrow (BM) by heavy chain (HC) VDJ recombination and expression of IgM associated with a surrogate LC on the cell surface. In a second round of recombi- nation, one o

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