cellulase and 2-phenyloxazolone

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.

Topics: Amino Acid Sequence; Animals; Base Sequence; Blotting, Western; Cellulase; Endopeptidases; Genetic Engineering; Genetic Vectors; Immunoglobulin Fab Fragments; Immunoglobulin G; Mice; Molecular Sequence Data; Oxazolone; Plasmids; Recombinant Fusion Proteins; Transfection; Trichoderma