Journal ArticleDOI
Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli
TLDR
Isolated variable domains may offer an alternative to monoclonal antibodies and serve as the key to building high-affinity human antibodies and the name 'single domain antibodies (dAbs)' is suggested for these antigen binding demands.Abstract:
IN antibodies, a heavy and a light chain variable domain, VH and VL, respectively, pack together and the hypervariable loops on each domain contribute to binding antigen1–4. We find, however, that isolated VH domains with good antigen-binding affinities can also be prepared. Using the polymerase chain reaction5, diverse libraries of VH genes were cloned from the spleen genomic DNA of mice immunized with either lysozyme or keyhole-limpet haemocyanin. From these libraries, VH domains were expressed and secreted from Escherichia coli. Binding activities were detected against both antigens, and two VH domains were characterized with affinities for lysozyme in the 20 nM range. Isolated variable domains may offer an alternative to monoclonal antibodies and serve as the key to building high-affinity human antibodies. We suggest the name 'single domain antibodies (dAbs)' for these antigen binding demands.read more
Citations
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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
Naturally occurring antibodies devoid of light chains
Cécile Hamers-Casterman,Touriya Atarhouch,Serge Muyldermans,Gene E. Robinson,Hamers C,Songa Eb,Bendahman N,Raymond Hamers +7 more
TL;DR: The presence of considerable amounts of IgG-like material of Mr 100K in the serum of the camel, which is composed of heavy-chain dimers and devoid of light chains, but nevertheless have an extensive antigen-binding repertoire, opens new perspectives in the engineering of antibodies.
Patent
Methods for producing members of specific binding pairs
John Mccafferty,Anthony Richard Pope,Kevin Stuart Johnson,Hoogenboom Hendricus R J M,Andrew D. Griffiths,Ronald Henry Jackson,Kaspar Philipp Holliger,James D. Marks,Timothy Piers Clackson,David John Chiswell,Gregory Paul Winter,Timothy Peter Seattle Bonnert +11 more
TL;DR: In this paper, a member of a specific binding pair (sbp) is identified by expressing DNA encoding a genetically diverse population of such sbp members in recombinant host cells in which the sbps members are displayed in functional form at the surface of a secreted recombinant genetic display package (rgdp) containing DNA encoding the sbp member or a polypeptide component thereof.
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.
Patent
Production of chimeric antibodies - a combinatorial approach
Hendricus Renerus Jacobus Matteus Hoogenboom,Michael Baier,Laurent Stephane Anne Therese Jespers,Gregory Paul Winter +3 more
TL;DR: In this paper, the authors describe methods for the production of antibodies, or antibody fragments, which have the same binding specificity as a parent antibody, but which have increased human characteristics.
References
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Journal ArticleDOI
Expression of an antibody Fv fragment in myeloma cells.
TL;DR: A heterodimer of both variable domains (Fv fragment), incorporating loops from an anti-lysozyme antibody, was expressed and secreted from myeloma cells in good yield, and shown to bind lysozyme.
Journal ArticleDOI
A homologue of the nuclear coded 49 kd subunit of bovine mitochondrial NADH-ubiquinone reductase is coded in chloroplast DNA.
TL;DR: The N‐terminal sequence of this protein has been determined and this has been used to design two mixtures of synthetic oligonucleotides, each containing 32 different sequences 17 bases long, which have been used as hybridization probes to isolate cDNA clones from a bovine library.
Journal ArticleDOI
A search for site-filling ligands in the Mcg Bence-Jones dimer: crystal binding studies of fluorescent compounds.
TL;DR: The dimer of the 6-isomer of carboxytetramethylrhodamine, in which the two carboxyl groups are in para positions on the phenyl moiety, proved to be an effective site-filling ligand and led to an explanation for isomeric discrimination in the binding site.
Journal ArticleDOI
Generation of a Catalytic Antibody by Site-Directed Mutagenesis
TL;DR: A hybrid Fv fragment of the dinitrophenyl-binding immunoglobulin A, MPOC315, has been generated by reconstituting a recombinant variable light chain produced in Escherichia coli with a variable heavy chain derived from the antibody.
Journal ArticleDOI
Overproduction of phage lambda repressor under control of the lac promotor of Escherichia coli.
TL;DR: The gene coding for bacteriophage Lambda repressor (cI gene) has been fused to the lac operon of Escherichia coli and in some of the fusions LambdaRepressor synthesis can be controlled by the lac operator and promoter.
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