title: Neutralization of Burkholderia pseudomallei protease by Fabs generated through phage display.
abstract: The isolation of therapeutic and functional protease inhibitors in vitro via combinatorial chemistry and phage display technology has been described previously. Here we report the construction of a combinatorial mouse-human chimeric antibody fragment (Fab) antibody library targeted against the protease of the tropical pathogen, Burkholderia pseudomallei. The resulting library was biopanned against the protease, and selected clones were analyzed for their ability to function as protease inhibitors. Three families of Fabs were identified by restriction fingerprinting, all of which demonstrated high specificity towards the protease of B. pseudomallei. Purified Fabs also demonstrated the capacity to inhibit B. pseudomallei protease activity in vitro, and this inhibitory property was exclusive to the pathogenic protease. Thus these recombinant antibodies are candidates for immunotherapy and tools to aid in further elucidation of the mechanism of action of the B. pseudomallei protease.
authors.affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Selanger.
authors.forename: Sheila
authors.initials: S
authors.lastname: Nathan
authors.forename: Christoph
authors.initials: C
authors.lastname: Rader
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Barbas
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016957:Burkholderia pseudomallei / Q000201:enzymology*; D016172:DNA Fingerprinting; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000494:pharmacology*; D051379:Mice; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000276:immunology; D019151:Peptide Library; D011480:Protease Inhibitors; D012313:RNA / Q000032:analysis / Q000096:biosynthesis; D013234:Stem Cells
chemical_list: D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011480:Protease Inhibitors; C034477:RNA primers; D012313:RNA; D010447:Peptide Hydrolases