Expression, engineering and characterization of the tumor-targeting heterodimeric immunocytokine F8-IL12.
Proinflammatory cytokines have been used for several years in patients with advanced cancer but their administration is typically associated with severe toxicity hampering their application to therapeutically active regimens. This problem can be overcome by using immunocytokines (cytokines fused to antibody or antibody fragments) which selectively deliver the active cytokine to the tumor environment. Preclinical and recent clinical results confirmed that this approach is a very promising avenue to go. We designed an immunocytokine consisting of the scFv(F8) specific to extra-domain A of fibronectin and the very potent human cytokine interleukin-12 (IL12). The heterodimeric nature of IL12 allows the engineering of various immunocytokine formats, based on different combinations of the two subunits (p35 and p40) together with the scFv. In comparison to monomeric or homodimeric cytokines, the construction of a heterodimeric immunocytokine poses many challenges, e.g. gene dosing, stable high-yield expression as well as good manufacture practice (GMP) purification and characterization. In this paper, we describe the successful construction, characterization and production of the heterodimeric immunocytokine F8-IL12. The positive outcome of this feasibility study leads now to GMP production of F8-IL12, which will soon enter clinical trials.
Sommavilla
Roberto
R
Philochem AG, c/o ETH Zürich, Institute of Pharmaceutical Sciences, Wolfgang-Pauli-Strasse 10, HCI E520, CH-8093 Zürich, Switzerland.
Pasche
Nadine
N
Trachsel
Eveline
E
Giovannoni
Leonardo
L
Roesli
Christoph
C
Villa
Alessandra
A
Neri
Dario
D
Kaspar
Manuela
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D061067:Antibodies, Monoclonal, Humanized; D001345:Autoradiography; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D053765:Interleukin-12 Subunit p35 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D053762:Interleukin-12 Subunit p40 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014018:Tissue Distribution; D014162:Transfection
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; C528643:F8 monoclonal antibody; D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies