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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A critical role for interleukin-1 receptor accessory protein in interleukin-1 signaling
TL;DR: It is shown that the recently identified IL‐1RI accessory protein (IL‐1RAcP) converts the silent into a fully functional IL-1RI complex, which is a critical early step in the signaling cascade mediated by the IL‐ 1RI activation complex.
Journal ArticleDOI
Identification of human antibody fragment clones specific for tetanus toxoid in a bacteriophage lambda immunoexpression library.
R L Mullinax,E A Gross,J R Amberg,B N Hay,H H Hogrefe,M M Kubitz,A Greener,M Alting-Mees,D Ardourel,Jay M. Short +9 more
TL;DR: A molecular biology approach to the identification of human monoclonal antibodies was applied and it is estimated that this human immunoexpression library contains 20,000 clones with high affinity and specificity to the authors' chosen antigen.
Patent
Methods for producing antibodies
TL;DR: In this article, a method of obtaining an antibody with a desired function was proposed, the method comprising: bringing a population of B cells into contact with a capturing agent, separating the captured B cells from the uncaptured B cells, culturing a plurality of captured cells wherein said B cells have not been sorted into single B cells immediately prior to culturing, screening the captured cells to identify cells capable of producing an antibody, and obtaining the desired antibody therefrom.
Journal ArticleDOI
Single chain Fab (scFab) fragment
Michael Hust,Thomas Jostock,Thomas Jostock,Christian Menzel,Christian Menzel,Bernd Voedisch,Anja Mohr,Anja Mohr,Mariam Brenneis,Mariam Brenneis,Martina Inga Kirsch,Doris Meier,Stefan Dübel +12 more
TL;DR: It is demonstrated that the introduction of a polypeptide linker between the fragment difficult (Fd) and the light chain (LC), resulting in the formation of a single chain Fab fragment (scFab), can lead to improved production of functional molecules.
Journal ArticleDOI
Crystal structure of HEL4, a soluble, refoldable human V(H) single domain with a germ-line scaffold.
TL;DR: The discovery and characterisation of an isolated human V(H) domain (HEL4) with properties similar to those of V(HH) domains are described, and the importance of a hydrophilic V(h):V(L) interface for purification of isolated V( H) domains is confirmed.
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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