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. 1999 Jun;80(8):1214–1222. doi: 10.1038/sj.bjc.6690488

An anti-CD30 single-chain Fv selected by phage display and fused to Pseudomonas exotoxin A (Ki-4(scFv)-ETÁ) is a potent immunotoxin against a Hodgkin-derived cell line

A Klimka 1,2, S Barth 1, B Matthey 1, R C Roovers 2, H Lemke 3, H Hansen 3, J-W Arends 2, V Diehl 1, H R Hoogenboom 2, A Engert 1
PMCID: PMC2362362  PMID: 10376974

Abstract

The human CD30 receptor is highly overexpressed on the surface of Hodgkin Reed-Sternberg cells and has been shown to be an excellent target for selective immunotherapy using monoclonal antibody-based agents such as immunotoxins. To construct a new recombinant immunotoxin for possible clinical use in patients with Hodgkin's lymphoma, we have chosen the murine anti-CD30 hybridoma Ki-4 to generate a high-affinity Ki-4 single-chain variable fragment (scFv). Hybridoma V-genes were polymerase chain reaction-amplified, assembled, cloned and expressed as a mini-library for display on filamentous phage. Functional Ki-4 scFv were obtained by selection of binding phage on the Hodgkin lymphoma-derived, CD30-expressing cell line L540Cy. The selected recombinant Ki-4 scFv was shown to specifically bind to an overlapping epitope on the CD30 antigen with binding kinetics similar to those of the original antibody. The Ki-4 scFv was subsequently fused to a deletion mutant of Pseudomonas exotoxin A (ETÁ). The resulting immunotoxin Ki-4(scFv)-ETÁ specifically binds to CD30+ L540Cy cells and inhibits the protein synthesis by 50% at a concentration (IC50) of 43 pM. This recombinant immunotoxin is a promising candidate for further clinical evaluation in patients with Hodgkin's lymphoma or other CD30+ malignancies. © 1999 Cancer Research Campaign

Keywords: CD30, Ki-4, phage display, recombinant immunotoxin, Hodgkin's lymphoma

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Footnotes

A Klimka and S Barth contributed equally to the work.

Selected References

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