Design of a human synthetic combinatorial library of single-chain antibodies.
Antibody libraries came into existence 15 years ago when the accumulating sequence data of immunoglobulin genes and the advent of the PCR technology made it possible to clone antibody gene repertoires. Phage display (most common) and additional display and screening technologies were applied to pan out desired binding specificities from antibody libraries. "Synthetic" or "semi-synthetic" libraries are from naïve, non-immunized source and considered to be a source for many different targets, including self-antigens. We describe here how to construct a large human synthetic single-chain Fv (scFv) antibody library displayed on phages, where in vivo-formed complementarity-determining regions (CDRs) are shuffled combinatorially onto germline-derived human variable-region frameworks.
Nahary
Limor
L
Tel-Aviv University, Ramat Aviv, Israel.
Benhar
Itai
I
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D000941:Antigens / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D022801:Complementarity Determining Regions / Q000276:immunology; D018274:Electroporation; D004926:Escherichia coli / Q000166:cytology; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D055767:Immobilized Proteins / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction
D000906:Antibodies; D000941:Antigens; D022801:Complementarity Determining Regions; D055767:Immobilized Proteins; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D019151:Peptide Library