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
Engineered antibody fragments and the rise of single domains
Philipp Holliger,Peter J Hudson +1 more
TL;DR: There is tremendous potential for all antibody fragments either as robust diagnostic reagents, or as nonimmunogenic in vivo biopharmaceuticals with superior biodistribution and blood clearance properties.
Journal ArticleDOI
Making antibody fragments using phage display libraries
TL;DR: Using a random combinatorial library of the rearranged heavy and kappa light chains from mice immune to the hapten 2-phenyloxazol-5-one (phOx), diverse libraries of antibody fragments are displayed on the surface of fd phage and elicited many more pairings with strong binding activities.
Journal ArticleDOI
Making Antibodies by Phage Display Technology
TL;DR: Human antibody fragments with many different binding specificities have been isolated from the same phage repertoire, including haptens, carbohydrates, secreted and cell surface proteins, viral coat proteins, and intracellular antigens from the lumen of the endoplasmic reticulum and the nucleus.
Journal ArticleDOI
Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda
William D. Huse,Lakshmi Sastry,Sheila A. Iverson,Angray S. Kang,Angray S. Kang,Michelle A. Alting-Mees,Dennis R. Burton,Dennis R. Burton,Stephen J. Benkovic,Stephen J. Benkovic,Richard A. Lerner +10 more
TL;DR: A novel bacteriophage lambda vector system was used to express in Escherichia coli a combinatorial library of Fab fragments of the mouse antibody repertoire, which allows rapid and easy identification of monoclonal Fab fragments in a form suitable for genetic manipulation.
Journal ArticleDOI
"Diabodies": small bivalent and bispecific antibody fragments
TL;DR: The design of small antibody fragments with two antigen-binding sites, which comprise a heavy-chain variable domain connected to a light- Chain variable domain on the same polypeptide chain (VH-VL), are described.
References
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Journal ArticleDOI
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Journal Article
Cleavage of structural proteins during the assemble of the head of bacterio-phage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
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Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
TL;DR: A method has been devised for the electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets that results in quantitative transfer of ribosomal proteins from gels containing urea.
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Experiments in molecular genetics
TL;DR: Molecular Genetics (Biology): An Overview | Sciencing Experimental in Molecular Genetics Experiments in molecular genetics (1972 edition) | Open ...
Journal ArticleDOI
Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors
TL;DR: New Escherichia coli host strains have been constructed for the E. coli bacteriophage M13 and the high-copy-number pUC-plasmid cloning vectors and mutations introduced into these strains improve cloning of unmodified DNA and of repetitive sequences.
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