title: Generation of human scFv antibody libraries: PCR amplification and assembly of light- and heavy-chain coding sequences.
abstract: The development of therapeutic antibodies for use in the treatment of human diseases has long been a goal for many researchers in the antibody field. One way to obtain these antibodies is through phage-display libraries constructed from human lymphocytes. This protocol describes the construction of human scFv (single chain antibody fragment) libraries using a short linker (GGSSRSS) or a long linker (GGSSRSSSSGGGGSGGGG). In this method, the individual rearranged heavy- and light-chain variable regions are amplified separately and are linked through a series of overlap polymerase chain reaction (PCR) steps to give the final scFv products that are used for cloning.
authors: Jennifer
authors: J
authors: Andris-Widhopf
authors: Peter
authors: P
authors: Steinberger
authors: Roberta
authors: R
authors: Fuller
authors: Christoph
authors: C
authors: Rader
authors: Carlos F
authors: CF
authors: Barbas
mesh_terms: D001688:Biological Products; D015723:Gene Library; D006801:Humans; D007155:Immunologic Factors / Q000235:genetics; D007158:Immunologic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics; D057127:Single-Chain Antibodies / Q000235:genetics*; D013678:Technology, Pharmaceutical / Q000379:methods*
chemical_list: D001688:Biological Products; D007155:Immunologic Factors; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies