title: Synthesis of high avidity antibody fragments (scFv multimers) for cancer imaging.
abstract: Multivalent antibody fragments (scFv dimers, trimers and tetramers) provide high avidity and ideal pharmacokinetics for tumour targeting applications. This protocol describes our optimised protocol for high-level bacterial synthesis of soluble antibody scFv fragments, as either monomers or multimers, using the heat-inducible bacterial expression vector pPOW3. Our protocol is rapid, which minimizes protein degradation, and utilises inexpensive reagents for cost-effective product synthesis. The strong, temperature-regulated promoters in pPOW3 provide efficient production of either monomeric or multimeric single-chain antibody fragments as dictated by the gene construct design.
authors.affiliation: CSIRO Health Sciences and Nutrition, 343 Royal Parade, Parkville, Victoria 3052, Australia. barbara.power@hsn.csiro.au
authors.forename: B E
authors.initials: BE
authors.lastname: Power
authors.forename: P J
authors.initials: PJ
authors.lastname: Hudson
mesh_terms: D000595:Amino Acid Sequence; D000912:Antibodies, Neoplasm / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000915:Antibody Affinity; D001483:Base Sequence; D003952:Diagnostic Imaging; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000276:immunology*; D013696:Temperature
chemical_list: D000912:Antibodies, Neoplasm; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; C055966:immunoglobulin Fv