Journal ArticleDOI
A directory of human germ-line Vχ segments reveals a strong bias in their usage
TLDR
Comparison with 236 rearranged sequences revealed that no more than 24 of these germ‐line sequences could be assigned rearranged counterparts, that some of these were rarely used, and that only about 11 sequences are used frequently, suggesting that the expressed Vχ repertoire is mainly derived from a limited number of segments.Abstract:
From the genomic DNA of a single individual, we have amplified, cloned and sequenced 37 human germ-line V kappa segments. Four of these segments were new. We then compiled a comprehensive directory of all germ-line V kappa segments and identified 50 different sequences with open reading frames. Comparison with 236 rearranged sequences revealed that no more than 24 of these germ-line sequences could be assigned rearranged counterparts, that some of these were rarely used, and that only about 11 sequences are used frequently. This suggests that the expressed V kappa repertoire is mainly derived from a limited number of segments. Most surprisingly, the J kappa-distal region of the locus appears to be rarely used: we could unambiguously assign 162 rearranged sequences to V kappa segments of the J kappa-proximal region, but only 5 to segments of the J kappa-distal region.read more
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Patent
Human antibodies derived from immunized xenomice
TL;DR: In this paper, a transgenic animal has been modified to produce antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, and various subsequent manipulations can be performed to obtain either antibodies per se or analogs thereof.
Journal ArticleDOI
Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library.
Tristan J. Vaughan,Andrew James Williams,Kevin Pritchard,Jane K. Osbourn,Anthony Richard Pope,John C. Earnshaw,John McCafferty,Regina A. Hodits,Jane Wilton,Kevin Stuart Johnson +9 more
TL;DR: This work shows that conventional hybridoma technology may be superseded by large phage libraries that are proving to be a stable and reliable source of specific, high affinity human monoclonal antibodies.
Journal ArticleDOI
Isolation of high affinity human antibodies directly from large synthetic repertoires.
Andrew D. Griffiths,S C Williams,Oliver Hartley,I M Tomlinson,Peter M. Waterhouse,William L. Crosby,Roland E. Kontermann,Peter T. Jones,N. M. Low,T. J. Allison +9 more
TL;DR: This work created highly diverse repertoires of heavy and light chains entirely in vitro from a bank of human V gene segments and generated a large synthetic repertoire of Fab fragments displayed on filamentous phage to help dissect the contributions of biological mechanisms and structural features governing V gene usage in vivo.
Patent
Transgenic mammals having human Ig loci including plural VH and VK regions and antibodies produced therefrom
TL;DR: In this paper, a transgenic non-human animal was engineered to contain human immunoglobulin gene loci, including plural variable (V H and Vκ) gene regions.
Journal ArticleDOI
Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides.
Achim Knappik,Liming Ge,Annemarie Honegger,Peter Pack,Melanie Fischer,Günter Wellnhofer,Adolf Hoess,Joachim Wölle,Andreas Plückthun,Bernhard Virnekäs +9 more
TL;DR: The small number of 49 master genes will allow future improvements to be incorporated quickly, and the separation of the frameworks may help in analyzing why nature has evolved these distinct subfamilies of antibody germline genes.
References
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Journal ArticleDOI
DNA sequencing with chain-terminating inhibitors
TL;DR: A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
Book
Experiments in molecular genetics
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Journal ArticleDOI
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.
Randall Keichi Saiki,Stephen J. Scharf,Fred A. Faloona,Kary B. Mullis,Glenn Thomas Horn,Henry A. Erlich,Norman Arnheim +6 more
TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
Journal ArticleDOI
Phage antibodies: filamentous phage displaying antibody variable domains
TL;DR: It is shown that complete antibody V domains can be displayed on the surface of fd bacteriophage, that the phage bind specifically to antigen and that rare phage can be isolated after affinity chromatography.
Journal ArticleDOI
By-passing immunization: Human antibodies from V-gene libraries displayed on phage
James D. Marks,Hennie R. Hoogenboom,Timothy Peter Bonnert,John McCafferty,Andrew D. Griffiths,Greg Winter +5 more
TL;DR: The results suggest that a single large phage display library can be used to isolate human antibodies against any antigen, by-passing both hybridoma technology and immunization.
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