Antibody-drug conjugates (ADCs) represent a promising class of biopharmaceuticals with the potential to localize at tumor site and improve therapeutic index cytotoxic drugs. While it is generally believed that ADCs need be internalized into cells in order display optimal activity, has recently been shown non-internalizing antibodies can efficiently liberate disulfide-linked drugs extracellular site, leading potent anti-cancer activity preclinical animal models. Here, we show engineered...

We describe the cloning and characterization of a novel fusion protein (termed L19‐mIL12), consisting murine interleukin‐12 in single‐chain format, sequentially fused to L19 antibody tandem diabody format. The bound avidly cognate antigen (the alternatively spliced EDB domain fibronectin), retained activity parental cytokine was able selectively localize tumors vivo , as shown by quantitative biodistribution analysis. L19‐mIL12 exhibited potent antitumor immunocompetent mice bearing CT26...

Abstract Purpose: There is a growing interest in the use of tumor antigens for therapeutic vaccination strategies. Unfortunately, most cases, peptide vaccines patients does not mediate shrinkage solid masses. Experimental Design: Here, we studied opportunity to boost with F8-TNF, an antibody fusion protein that selectively delivers TNF extracellular matrix. AH1, model antigen investigate CD8+ T-cell immunity BALB/c mice, was used as vaccine. Results: Peptide alone exhibited only modest...

Engineered cytokines are gaining importance in cancer therapy, but these products often limited by toxicity, especially at early time points after intravenous administration. 4-1BB is a member of the tumor necrosis factor receptor superfamily, which has been considered as target for therapeutic strategies with agonistic antibodies or using its cognate cytokine ligand, 4-1BBL. Here we describe engineering an antibody fusion protein, termed F8-4-1BBL, that does not exhibit activity solution...

Mice bearing CT26 tumors can be cured by administration of L19-mIL12 or F8-mTNF, two antibody fusion proteins which selectively deliver their cytokine payload to the tumor. In both settings, cancer cures crucially depended on CD8+ T cells and AH1 peptide (derived from gp70 protein murine leukemia virus) acted as main tumor-rejection antigen, with ∼50% in neoplastic mass being AH1-specific after therapy. order characterize clonality cell response, its phenotype, activation status, we isolated...

Abstract IL15 is an immunostimulatory cytokine that holds promises for cancer therapy, but its performance (alone or as partner fusion proteins) has often been limited by suboptimal accumulation in the tumor and very rapid clearance from circulation. Most recently, Sushi Domain (SD, shortest region of receptor α, capable binding to IL15) fused IL15-based anticancer products increase biological activity. Here, we describe two novel antibody proteins (termed F8-F8-IL15 F8-F8-SD-IL15), specific...

There is a growing interest in the use of patient-derived T cells for treatment various types malignancies. The expansion polyclonal and polyspecific population tumor-reactive cells, with subsequent infusion into same donor patient, has been implemented, sometimes positive results. It not known, however, whether set single antigen specificity may be sufficient an effective therapy. To gain more insights this matter, we used naturally occurring recognizing retroviral peptide (AH1), which...

Antibody-cytokine fusion proteins can have the potential to increase density and activity of subsets leukocytes within tumor mass. Here, we describe design, production, characterization four novel antibody-cytokine directed against human carbonic anhydrase IX, a highly validated marker hypoxia that is overexpressed in clear cell renal carcinoma other malignancies. As immunomodulatory payloads used TNF, IL2, IFNα2 (corresponding products are clinical use), IL12 (as this cytokine potently...

The targeted delivery of interleukin-2 to the tumor is gaining attention as an avenue potentiate action T and NK cells at site disease. We have previously described fusion L19 antibody, specific EDB domain fibronectin, with human interleukin-2, using a non-covalent homodimeric diabody format. Here, we describe four novel formats for L19-IL2 fusion, featuring different arrangements antibody IL2. A comparative quantitative biodistribution analysis in tumor-bearing mice radioiodinated proteins...

// Michael R. Mortensen 1 , Jacqueline Mock Marco Bertolini Stringhini Catalano and Dario Neri Department of Chemistry Applied Biosciences (D-CHAB), Institute for Pharmaceutical Sciences (IPW), 8093 Zurich, Switzerland Correspondence to: Mortensen, email: michael.mortensen@pharma.ethz.ch Keywords: immunotherapy; immunocytokines; EDB fibronectin; antibody-cytokine fusion proteins; engineered cytokine Received: July 30, 2020 Accepted: September 24, Published: November 03, Copyright: © et al....

LIGHT is a member of the tumor necrosis factor superfamily, which has been claimed to mediate anti-tumor activity on basis cancer cures observed in immunocompetent mice bearing transgenic LIGHT-expressing tumors. The preclinical development LIGHT-based therapeutic hindered by lack functional stability exhibited this protein. Here, we describe cloning, expression, and characterization five antibody-LIGHT fusion proteins, directed against alternatively spliced extra domain A fibronectin,...

Abstract The possibility to cure immunocompetent mice bearing murine CT26 colorectal tumors using cytokine-based therapeutics allows study the tumor rejection process at a molecular level. Following treatment with L19-mIL12 or F8-mTNF, two antibody fusion proteins which preferentially concentrate cytokine payload site, could be cured in that crucially relies on CD8+ T cells. In both settings, AH1 peptide (derived from gp70 envelop protein of leukemia virus) acted as main antigen and ~50%...

ABSTRACT We describe the cloning and characterization of a novel fusion protein (termed L19-mIL12), consisting murine interleukin-12 in single-chain format, sequentially fused to L19 anti-body tandem diabody format. The bound avidly cognate antigen (the alternatively-spliced EDB domain fibronectin), retained activity parental cyto-kine was able selectively localize tumors vivo , as shown by quantitative biodistribution analysis. L19-mIL12 exhibited potent anti-tumor immunocompetent mice...

ABSTRACT Engineered cytokines are gaining importance for cancer therapy but those products often limited by toxicity, especially at early time points after intravenous administration. 4-1BB is a member of the tumor necrosis factor receptor superfamily, which has been considered as target therapeutic strategies with agonistic antibodies or using its cognate cytokine ligand, 4-1BBL. Here we describe engineering an antibody fusion protein (termed F8-4-1BBL), does not exhibit activity in...

<p>Analysis of AH1-specific CD8+ T cells in WEHI-164 tumor-bearing mice</p>

<p>MaxQuant raw output, including peptide sequence and modifications, precursor mass charge as well scores associated statistical information</p>

<div>AbstractPurpose:<p>There is a growing interest in the use of tumor antigens for therapeutic vaccination strategies. Unfortunately, most cases, peptide vaccines patients does not mediate shrinkage solid masses.</p><p><b>Experimental Design:</b> Here, we studied opportunity to boost with F8-TNF, an antibody fusion protein that selectively delivers TNF extracellular matrix. AH1, model antigen investigate CD8<sup>+</sup> T-cell immunity BALB/c...

<div>AbstractPurpose:<p>There is a growing interest in the use of tumor antigens for therapeutic vaccination strategies. Unfortunately, most cases, peptide vaccines patients does not mediate shrinkage solid masses.</p><p><b>Experimental Design:</b> Here, we studied opportunity to boost with F8-TNF, an antibody fusion protein that selectively delivers TNF extracellular matrix. AH1, model antigen investigate CD8<sup>+</sup> T-cell immunity BALB/c...

<p>Toxicity data of the therapy studies</p>

<p>Sequences from gp70 and results the scoring against H2 I-Ad I-Ed position-specific functions</p>

<p>MaxQuant raw output, including peptide sequence and modifications, precursor mass charge as well scores associated statistical information</p>

<p>Toxicity data of the therapy studies</p>

<p>Analysis of AH1-specific CD8+ T cells in WEHI-164 tumor-bearing mice</p>