Nyyssönen E, Penttilä M, Harkki A, Saloheimo A, Knowles J K, Keränen S
VTT Biotechnical Laboratory, Espoo, Finland.
Biotechnology (N Y). 1993 May;11(5):591-5. doi: 10.1038/nbt0593-591.
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.
我们对丝状真菌里氏木霉进行了改造，使其能够在培养基中组装并分泌具有免疫原性的工程化Fab抗体片段。通过将重链Fd与里氏木霉纤维素酶CBHI融合，产量得到了显著提高。分泌的具有免疫活性的Fab和CBHI-Fab融合蛋白的产量分别为1mg/L和150mg/L。Fab片段可通过里氏木霉的一种胞外蛋白酶从融合蛋白CBHI-Fab中释放出来。工程化Fab与独特型抗体之间在对抗原的亲和力上没有可检测到的差异。
