Efficient production of antibody fragments by the filamentous fungus Trichoderma reesei.
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.
Nyyssönen
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E
VTT Biotechnical Laboratory, Espoo, Finland.
Penttilä
M
M
Harkki
A
A
Saloheimo
A
A
Knowles
J K
JK
Keränen
S
S
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
D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; C018361:2-phenyloxazolone; D010081:Oxazolone; D002480:Cellulase; D010450:Endopeptidases