Cloning and characterization of novel tumor-targeting immunocytokines based on murine IL7

Nadine Pasche, Janine Woytschak, Sarah Wulhfard, Alessandra Villa, Katharina Frey, Dario Neri

Research output: Contribution to journalArticle

Abstract

We generated and characterized novel antibody-cytokine fusion proteins (" immunocytokines") based on murine interleukin-7 (IL7), an immunomodulatory protein which has previously shown anti-cancer activity in preclinical models and whose human counterpart is currently being investigated in clinical trials. The sequential fusion of the clinical-stage antibody fragment scFv(F8), specific to a tumor-associated splice isoform of fibronectin, yielded an immunocytokine (termed "F8-mIL7") of insufficient pharmaceutical quality and in vivo tumor targeting performance, with a striking dose dependence on tumor targeting selectivity. By contrast, a novel immunocytokine design (termed "F8-mIL7-F8"), in which two scFv moieties were fused at the N- and C-terminus of murine IL7, yielded a protein of excellent pharmaceutical quality and with improved tumor-targeting performance [tumor: blood ratio. = 16:1, 24 h after injection]. Both F8-mIL7 and F8-mIL7-F8 could induce tumor growth retardation in immunocompetent mice, but were not able to eradicate F9 tumors. The combination of F8-mIL7-F8 with paclitaxel led to improved therapeutic results, which were significantly better compared to those obtained with saline treatment. The study indicates how the engineering of novel immunocytokine formats may help generate fusion proteins of acceptable pharmaceutical quality, for those immunomodulatory proteins which do not lend themselves to a direct fusion with antibody fragments.

Original languageEnglish
Pages (from-to)84-92
Number of pages9
JournalJournal of Biotechnology
Volume154
Issue number1
DOIs
Publication statusPublished - Jun 10 2011

Fingerprint

Interleukin-7
Cloning
Organism Cloning
Tumors
Proteins
Fusion reactions
Neoplasms
Antibodies
Drug products
Immunoglobulin Fragments
Pharmaceutical Preparations
Paclitaxel
Fibronectins
Protein Isoforms
Blood
Cytokines
Clinical Trials
Injections
Therapeutics
Growth

Keywords

  • F8 antibody
  • Immunocytokine
  • Interleukin 7
  • Targeting

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Cloning and characterization of novel tumor-targeting immunocytokines based on murine IL7. / Pasche, Nadine; Woytschak, Janine; Wulhfard, Sarah; Villa, Alessandra; Frey, Katharina; Neri, Dario.

In: Journal of Biotechnology, Vol. 154, No. 1, 10.06.2011, p. 84-92.

Research output: Contribution to journalArticle

Pasche, N, Woytschak, J, Wulhfard, S, Villa, A, Frey, K & Neri, D 2011, 'Cloning and characterization of novel tumor-targeting immunocytokines based on murine IL7', Journal of Biotechnology, vol. 154, no. 1, pp. 84-92. https://doi.org/10.1016/j.jbiotec.2011.04.003
Pasche, Nadine ; Woytschak, Janine ; Wulhfard, Sarah ; Villa, Alessandra ; Frey, Katharina ; Neri, Dario. / Cloning and characterization of novel tumor-targeting immunocytokines based on murine IL7. In: Journal of Biotechnology. 2011 ; Vol. 154, No. 1. pp. 84-92.
@article{296e06a49c3d496f88e5f47955aeddce,
title = "Cloning and characterization of novel tumor-targeting immunocytokines based on murine IL7",
abstract = "We generated and characterized novel antibody-cytokine fusion proteins ({"} immunocytokines{"}) based on murine interleukin-7 (IL7), an immunomodulatory protein which has previously shown anti-cancer activity in preclinical models and whose human counterpart is currently being investigated in clinical trials. The sequential fusion of the clinical-stage antibody fragment scFv(F8), specific to a tumor-associated splice isoform of fibronectin, yielded an immunocytokine (termed {"}F8-mIL7{"}) of insufficient pharmaceutical quality and in vivo tumor targeting performance, with a striking dose dependence on tumor targeting selectivity. By contrast, a novel immunocytokine design (termed {"}F8-mIL7-F8{"}), in which two scFv moieties were fused at the N- and C-terminus of murine IL7, yielded a protein of excellent pharmaceutical quality and with improved tumor-targeting performance [tumor: blood ratio. = 16:1, 24 h after injection]. Both F8-mIL7 and F8-mIL7-F8 could induce tumor growth retardation in immunocompetent mice, but were not able to eradicate F9 tumors. The combination of F8-mIL7-F8 with paclitaxel led to improved therapeutic results, which were significantly better compared to those obtained with saline treatment. The study indicates how the engineering of novel immunocytokine formats may help generate fusion proteins of acceptable pharmaceutical quality, for those immunomodulatory proteins which do not lend themselves to a direct fusion with antibody fragments.",
keywords = "F8 antibody, Immunocytokine, Interleukin 7, Targeting",
author = "Nadine Pasche and Janine Woytschak and Sarah Wulhfard and Alessandra Villa and Katharina Frey and Dario Neri",
year = "2011",
month = "6",
day = "10",
doi = "10.1016/j.jbiotec.2011.04.003",
language = "English",
volume = "154",
pages = "84--92",
journal = "Journal of Biotechnology",
issn = "0168-1656",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Cloning and characterization of novel tumor-targeting immunocytokines based on murine IL7

AU - Pasche, Nadine

AU - Woytschak, Janine

AU - Wulhfard, Sarah

AU - Villa, Alessandra

AU - Frey, Katharina

AU - Neri, Dario

PY - 2011/6/10

Y1 - 2011/6/10

N2 - We generated and characterized novel antibody-cytokine fusion proteins (" immunocytokines") based on murine interleukin-7 (IL7), an immunomodulatory protein which has previously shown anti-cancer activity in preclinical models and whose human counterpart is currently being investigated in clinical trials. The sequential fusion of the clinical-stage antibody fragment scFv(F8), specific to a tumor-associated splice isoform of fibronectin, yielded an immunocytokine (termed "F8-mIL7") of insufficient pharmaceutical quality and in vivo tumor targeting performance, with a striking dose dependence on tumor targeting selectivity. By contrast, a novel immunocytokine design (termed "F8-mIL7-F8"), in which two scFv moieties were fused at the N- and C-terminus of murine IL7, yielded a protein of excellent pharmaceutical quality and with improved tumor-targeting performance [tumor: blood ratio. = 16:1, 24 h after injection]. Both F8-mIL7 and F8-mIL7-F8 could induce tumor growth retardation in immunocompetent mice, but were not able to eradicate F9 tumors. The combination of F8-mIL7-F8 with paclitaxel led to improved therapeutic results, which were significantly better compared to those obtained with saline treatment. The study indicates how the engineering of novel immunocytokine formats may help generate fusion proteins of acceptable pharmaceutical quality, for those immunomodulatory proteins which do not lend themselves to a direct fusion with antibody fragments.

AB - We generated and characterized novel antibody-cytokine fusion proteins (" immunocytokines") based on murine interleukin-7 (IL7), an immunomodulatory protein which has previously shown anti-cancer activity in preclinical models and whose human counterpart is currently being investigated in clinical trials. The sequential fusion of the clinical-stage antibody fragment scFv(F8), specific to a tumor-associated splice isoform of fibronectin, yielded an immunocytokine (termed "F8-mIL7") of insufficient pharmaceutical quality and in vivo tumor targeting performance, with a striking dose dependence on tumor targeting selectivity. By contrast, a novel immunocytokine design (termed "F8-mIL7-F8"), in which two scFv moieties were fused at the N- and C-terminus of murine IL7, yielded a protein of excellent pharmaceutical quality and with improved tumor-targeting performance [tumor: blood ratio. = 16:1, 24 h after injection]. Both F8-mIL7 and F8-mIL7-F8 could induce tumor growth retardation in immunocompetent mice, but were not able to eradicate F9 tumors. The combination of F8-mIL7-F8 with paclitaxel led to improved therapeutic results, which were significantly better compared to those obtained with saline treatment. The study indicates how the engineering of novel immunocytokine formats may help generate fusion proteins of acceptable pharmaceutical quality, for those immunomodulatory proteins which do not lend themselves to a direct fusion with antibody fragments.

KW - F8 antibody

KW - Immunocytokine

KW - Interleukin 7

KW - Targeting

UR - http://www.scopus.com/inward/record.url?scp=79957437540&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79957437540&partnerID=8YFLogxK

U2 - 10.1016/j.jbiotec.2011.04.003

DO - 10.1016/j.jbiotec.2011.04.003

M3 - Article

C2 - 21527292

AN - SCOPUS:79957437540

VL - 154

SP - 84

EP - 92

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

IS - 1

ER -