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
More filters
Journal ArticleDOI
The structural basis of antibody-antigen recognition.
TL;DR: Beyond improving the understanding of immunity, characterization of the functional role of different parts of the Ab molecule may help in Ab engineering, design of CDR-derived peptides, and epitope prediction.
Patent
Production of antibodies using gene libraries
TL;DR: A method of producing libraries of genes encoding antigen-combining molecules or antibodies is described in this article. But this method does not require an in vivo procedure and is not suitable for the detection, quantification, purification and neutralization of antigens, as well as for diagnostic, therapeutic and prophylactic purposes.
Journal ArticleDOI
Recognition of antigens by single-domain antibody fragments: the superfluous luxury of paired domains
TL;DR: The mutations and structural adaptations that have taken place to reshape a VH of a Vh-VL pair into a single-domain VHH with retention of a sufficient variability are reviewed.
Book ChapterDOI
Protein engineering of single-chain Fv analogs and fusion proteins.
James S. Huston,Meredith Mudgett-Hunter,Mei-Sheng Tai,John Mccartney,Frederick Warren,Edgar Haber,Hermann Oppermann +6 more
TL;DR: This chapter describes the minimal antibody binding site through the protein engineering of single-chain Fv analog and fusion proteins giving insight into their design, production, and properties.
Journal ArticleDOI
Natural and designer binding sites made by phage display technology
TL;DR: This application of phage display technology is reviewed, and the many other antibody-engineering avenues this technology offers are highlighted.
References
More filters
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.
Journal ArticleDOI
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.
Book
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.
Related Papers (5)
Continuous cultures of fused cells secreting antibody of predefined specificity
G Köhler,C. Milstein +1 more