title: Efficient production of antibody fragments by the filamentous fungus Trichoderma reesei.
abstract: We have engineered the filamentous fungus Trichoderma reesei to assemble and secrete immunologically authentic engineered Fab antibody fragments into the culture medium. A major improvement in yield was achieved by fusing the heavy Fd chain to the T. reesei cellulase, CBHI. The yields of secreted, immunologically active Fab and CBHI-Fab fusion were 1 mg/l and 150 mg/l, respectively. The Fab fragment can be released from the fusion protein CBHI-Fab by an extracellular T. reesei protease. There was no detectable difference in affinity for the antigen between the engineered Fab and the idiotypic antibody.
authors.affiliation: VTT Biotechnical Laboratory, Espoo, Finland.
authors.forename: E
authors.initials: E
authors.lastname: Nyyssönen
authors.forename: M
authors.initials: M
authors.lastname: Penttilä
authors.forename: A
authors.initials: A
authors.lastname: Harkki
authors.forename: A
authors.initials: A
authors.lastname: Saloheimo
authors.forename: J K
authors.initials: JK
authors.lastname: Knowles
authors.forename: S
authors.initials: S
authors.lastname: Keränen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002480:Cellulase / Q000235:genetics; D010450:Endopeptidases / Q000378:metabolism; D005818:Genetic Engineering; D005822:Genetic Vectors; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010081:Oxazolone / Q000031:analogs & derivatives / Q000276:immunology; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D014242:Trichoderma / Q000235:genetics* / Q000276:immunology
chemical_list: D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; C018361:2-phenyloxazolone; D010081:Oxazolone; D002480:Cellulase; D010450:Endopeptidases