Evaluation of human pancreatic RNase as effector molecule in a therapeutic antibody platform

Human antibody-ribonuclease (RNase) fusion proteins, referred to as immunoRNases, have been proposed as an alternative to heterologous immunotoxins, without their immunogenicity and unspecific toxicity issues. In this study, we investigated if human pancreatic RNase will be suitable as effector comp...

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Veröffentlicht in:mAbs 2014-03, Vol.6 (2), p.367-380
Hauptverfasser: Schirrmann, Thomas, Frenzel, André, Linden, Lars, Stelte-Ludwig, Beatrix, Willuda, Jörg, Harrenga, Axel, Dübel, Stefan, Müller-Tiemann, Beate, Trautwein, Mark
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container_end_page 380
container_issue 2
container_start_page 367
container_title mAbs
container_volume 6
creator Schirrmann, Thomas
Frenzel, André
Linden, Lars
Stelte-Ludwig, Beatrix
Willuda, Jörg
Harrenga, Axel
Dübel, Stefan
Müller-Tiemann, Beate
Trautwein, Mark
description Human antibody-ribonuclease (RNase) fusion proteins, referred to as immunoRNases, have been proposed as an alternative to heterologous immunotoxins, without their immunogenicity and unspecific toxicity issues. In this study, we investigated if human pancreatic RNase will be suitable as effector component in a therapeutic antibody development platform. We generated several fusion proteins consisting of tumor-specific human immunoglobulins (IgGs) and human pancreatic RNase. Transient mammalian cell production was efficient and IgG-RNases were purified to homogeneity. Antigen binding was comparable to the parental antibodies and RNase catalytic activity was retained even in the presence of 50-fold molar excess of human cytosolic RNase inhibitor (RI). Serum stability, cell binding and internalization of IgG-RNases were comparable to the parental IgGs. Despite these promising properties, none of the IgG-RNases revealed significant inhibition of tumor cell growth in vitro even when targeting different antigens putatively employing different endocytotic pathways. The introduction of different linkers containing endosomal protease cleavage sites into the IgG-RNase did not enhance cytotoxicity. Similarly, RI evasive human pancreatic RNase variants mediated only small inhibiting effects on tumor cell growth at high concentrations, potentially reflecting inefficient cytosolic translocation. Taken together, human pancreatic RNase and variants did not prove to be generally suitable as effector component for a therapeutic antibody drug development platform.
doi_str_mv 10.4161/mabs.27830
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subjects Adenocarcinoma - drug therapy
Adenocarcinoma - immunology
antibodies
Antibodies, Catalytic - genetics
Antibodies, Catalytic - metabolism
Antigens, Neoplasm - immunology
cancer therapy
Cell Growth Processes - drug effects
Colonic Neoplasms - drug therapy
Colonic Neoplasms - immunology
Cytotoxicity, Immunologic
Endocytosis
HEK293 Cells
HT29 Cells
human pancreatic RNase
Humans
IgG
immunoglobulin
Immunoglobulin G - genetics
Immunoglobulin G - metabolism
immunoRNase
Immunotherapy - methods
Immunotherapy - trends
Lung Neoplasms - drug therapy
Lung Neoplasms - immunology
Molecular Targeted Therapy
Pancreas - enzymology
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Ribonucleases - genetics
Ribonucleases - metabolism
RNase inhibitor
title Evaluation of human pancreatic RNase as effector molecule in a therapeutic antibody platform
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