title: Cloning and expression of single chain antibody fragments in Escherichia coli and Pichia pastoris.
abstract: The potential of recombinant antibody fragments is likely to be fulfilled only if they can be produced routinely at high concentrations. We have compared the ability of Escherichia coli and Pichia pastoris to produce functional recombinant single chain antibody (scAb) fragments. Two scAb fragments were expressed, an antihuman type V acid phosphatase (TRAP) and an anti-Pseudomonas aeruginosa lipoprotein I. We report here that, while expression from P. pastoris resulted in a significantly increased level of expression of the anti-TRAP scAb compared to E. coli, neither fragment was able to bind its target antigen as well as the bacterial product.
authors.affiliation: Department of Molecular and Cell Biology, University of Aberdeen, Scotland.
authors.forename: P M
authors.initials: PM
authors.lastname: Cupit
authors.forename: J A
authors.initials: JA
authors.lastname: Whyte
authors.forename: A J
authors.initials: AJ
authors.lastname: Porter
authors.forename: M J
authors.initials: MJ
authors.lastname: Browne
authors.forename: S D
authors.initials: SD
authors.lastname: Holmes
authors.forename: W J
authors.initials: WJ
authors.lastname: Harris
authors.forename: C
authors.initials: C
authors.lastname: Cunningham
mesh_terms: D000135:Acid Phosphatase / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D005803:Genes, Immunoglobulin; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D008074:Lipoproteins / Q000276:immunology*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D014169:Transformation, Bacterial; D014170:Transformation, Genetic
chemical_list: D000941:Antigens; D001426:Bacterial Proteins; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D008074:Lipoproteins; D011994:Recombinant Proteins; C060507:outer membrane lipoprotein I, Bacteria; D000135:Acid Phosphatase