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
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An enediyne-energized single-domain antibody-containing fusion protein shows potent antitumor activity.
TL;DR: The results showed that energized fusion protein VH-LDP-AE was more effective than lidamycin and mitomycin and suggested that Vh-L DP-AE would be a promising candidate for cancer-targeting therapy.
Journal ArticleDOI
Production of single chain Fab (scFab) fragments in Bacillus megaterium
TL;DR: The Gram-positive bacterium Bacillus megaterium allows the secretory production of antibody fragments including the novel scFab antibody format and the yield and quality of functional antibody fragment is comparable to the periplasmic production in E. coli.
Patent
Prodrugs activated by targeted catalytic proteins
John H. Kenten,Reid Von Borstel,Jan Casadei,Balreddy Kamireddy,Mark T. Martin,Richard J. Massey,Andrew D. Napper,David M. Simpson,Rodger Smith,Richard C. Titmas,Williams Richard O +10 more
TL;DR: In this paper, the authors proposed a method to activate prodrugs activated by catalytic proteins, such as enzymes and catalytic antibodies, as cytotoxic chemotherapeutic agents.
Journal ArticleDOI
Anti-TNFα domain antibody construct CEP-37247: Full antibody functionality at half the size.
Adam Clarke,Zehra Elgundi,Teresa Domagala,Raina J. Simpson,Nga B. Le,Anthony Gerard Doyle,Phil A. Jennings +6 more
TL;DR: Preclinical data for CEP-37247 show that the bivalent anti-tumor necrosis factor (TNF)α domain antibody protein construct is capable of mediating antibody-dependent cell-mediated cytotoxicity and has a circulating half-life of approximately 4.5 days in cynomolgus macaques.
Journal ArticleDOI
Universal cloning and direct sequencing of rearranged antibody V genes using C region primers, biotin-captured cDNA and one-side PCR.
TL;DR: A rapid and 'universal' strategy for cloning rearranged antibody genes from any species for which the sequence of the C segment(s) are known and will find applications in three areas: cloning rearrange antibody genes in all cases in which cloning with V-J primers is not possible; repertoire studies in which an unbiased cloning procedure is required for accurate estimate of gene usage; and generation of VH and VL gene libraries from immunised animals.
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.
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.
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