Human angiogenin fused to human CD30 ligand (Ang-CD30L) exhibits specific cytotoxicity against CD30-positive lymphoma.

摘要: A number of different immunotoxins composed cell-specific targeting structures coupled to plant or bacterial toxins have increasingly been evaluated for immunotherapy. Because these foreign proteins are highly immunogenic in humans, we developed a new CD30 ligand-based fusion toxin (Ang-CD30L) using the human RNase angiogenin. The completely gene was inserted into pET-based expression plasmid. Transformed Escherichia coli BL21(DE3) were grown under osmotic stress conditions presence compatible solutes. After isopropyl β-d-thiogalactoside induction, M r 37,000 His 10 -tagged Ang-CD30L directed periplasmic space and functionally purified by combination metal ion affinity followed enterokinase cleavage -Tag molecular size chromatography. characteristics recombinant protein assessed ELISA, flow cytometry, toxicity assays showing specific activity against + Hodgkin-derived cells. Specific binding verified competition with anti-CD30 monoclonal antibody Ki-4 commercially available CD30L-CD8 chimeric protein. showed vitro . immunotoxin significant toward several CD30-positive cell lines (HDLM-2, L1236, KM-H2, L540Cy) exhibited highest cytotoxicity L540 cells (IC 50 = 8 ng/ml) as determined proliferation assays. specificity confirmed competitive This is first report on possibly largely reduced immunogenicity treatment malignancies.