Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Friedman, Mikaela; Lindström, Sara; Ekerljung, Lina; Andersson-Svahn, Helene; Carlsson, Jörgen; Brismar, Hjalmar; Gedda, Lars; Frejd, Fredrik Y.; Ståhl, Stefan

doi:10.1042/ba20090096

Cited by 60 publications

(33 citation statements)

References 46 publications

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“…One possible application for bispecific molecules would be to bind surface receptors on tumor cells in such a way that active dimerization and, thus, cell signaling, is sterically inhibited (29). Another attractive property for bispecific molecules is the capacity for binding two tumor-associated receptors to increase the selectivity by preferably directing the agents to cells expressing both receptors instead of binding to cells expressing only one of these receptors.…”

Section: Discussionmentioning

confidence: 99%

A Bispecific Enediyne-Energized Fusion Protein Containing Ligand-Based and Antibody-Based Oligopeptides against Epidermal Growth Factor Receptor and Human Epidermal Growth Factor Receptor 2 Shows Potent Antitumor Activity

Guo

Zhu

Shang

et al. 2010

Clinical Cancer Research

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Purpose: The cooverexpression of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) observed in many human tumors and their synergistic interaction in the transformation of cells make these receptors important targets for the development of new targeted therapeutics. Targeting of EGFR and HER2 simultaneously has been pursued as a strategy with which to potentially increase efficiency and selectivity in therapy of certain cancers. This study was set to construct a bispecific energized fusion protein (Ec-LDP-Hr-AE) consisting of two oligopeptides against EGFR and HER2, and lidamycin, and investigate its antitumor efficacy.Experimental Design: In vitro experiments measured the binding and internalization of bispecific Ec-LDP-Hr fusion protein. The potency of energized fusion proteins was also done in which the bispecific Ec-LDP-Hr-AE was compared with lidamycin (LDM) and its monospecific counterparts, Ec-LDP-AE and LDP-Hr-AE. In vivo, Ec-LDP-Hr-AE was given i.v. to nude mice bearing human ovarian carcinoma SK-OV-3 xenografts.Results: Binding and internalization studies showed that bispecific fusion protein Ec-LDP-Hr bound to carcinoma cells specifically and then were internalized into the cytoplasm. Bispecific Ec-LDP-Hr-AE was more potent and selective in its cytotoxicity against different carcinoma cell lines than corresponding momospecific agents and LDM in vitro. In addition, Ec-LDP-Hr-AE significantly inhibited the growth of SK-OV-3 xenografts in nude mouse model. In vivo imaging study showed that FITC-labeled Ec-LDP-Hr was targeted and accumulated in the tumors.Conclusion: A ligand-based and an antibody-based oligopeptide fused to the enediyne antibiotic LDM created a new bispecific fusion protein with low molecular weight and more potent in vitro and in vivo antitumor activity (than momospecific fusion proteins). Clin Cancer Res; 16(7); 2085-94. ©2010 AACR.The ErbB family (ErbB1/EGFR, ErbB2/HER2, ErbB3/ HER3, and ErbB4/HER4) of receptor tyrosine kinases is known to drive both formation and progression of several commonly occurring cancers (1). Ligand binding to the receptors results in receptor homodimerization and heterodimerization, then initiated a series of intracellular events that ultimately promote cell growth, proliferation, differentiation, and migration (2-4). Human epidermal growth factor receptor 2 (HER2) has no identified ligand but it is the preferred binding partner for the other three family members (5). The cooverexpression of epidermal growth factor receptor (EGFR) and HER2 observed in many human tumors and their synergistic interaction in the transformation of cells make these receptors important targets for the development of new targeted therapeutics (6-8). Several monoclonal antibodies (e.g., cetuximab, trastuzumab, and pertuzumab) and tyrosine kinase inhibitors (e.g., gefitinib, erlotinib, and lapatinib) targeting EGFR and HER2 or both have been approved by the U.S. Food and Drug Administration for therapeutic use and several more are in ...

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Section: Discussionmentioning

confidence: 99%

A Bispecific Enediyne-Energized Fusion Protein Containing Ligand-Based and Antibody-Based Oligopeptides against Epidermal Growth Factor Receptor and Human Epidermal Growth Factor Receptor 2 Shows Potent Antitumor Activity

Guo

Zhu

Shang

et al. 2010

Clinical Cancer Research

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“…This alternative to recombinant bacterial production enables site-specific incorporation of various chemical moieties, such as fluorescent probes or chelating groups for binding radioactive metal atoms in a single chemical process. Furthermore, the small size of the scaffold provides means for modular approaches, where different Affibody molecules are combined into fusion proteins with multiple properties while still retaining an overall size much smaller compared to a full-length antibody [13].…”

Section: Fusion Proteinsmentioning

confidence: 99%

Affibody molecules: Engineered proteins for therapeutic, diagnostic and biotechnological applications

et al. 2010

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a b s t r a c tAffibody molecules are a class of engineered affinity proteins with proven potential for therapeutic, diagnostic and biotechnological applications. Affibody molecules are small (6.5 kDa) single domain proteins that can be isolated for high affinity and specificity to any given protein target. Fifteen years after its discovery, the Affibody technology is gaining use in many groups as a tool for creating molecular specificity wherever a small, engineering compatible tool is warranted. Here we summarize recent results using this technology, propose an Affibody nomenclature and give an overview of different HER2-specific Affibody molecules. Cumulative evidence suggests that the three helical scaffold domain used as basis for these molecules is highly suited to create a molecular affinity handle for vastly different applications.

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“…Due to their simple structure sdAbs and antibody mimetics are very easy to manipulate, engineer and produce. It is even possible to fuse two single domains to generate bispecific molecules [112]. However, there are also certain drawbacks associated with these molecules.…”

Section: Evolution Of Bispecific Antibodiesmentioning

confidence: 99%

Cancer Immunotherapy by Retargeting of Immune Effector Cells via Recombinant Bispecific Antibody Constructs

Stamova¹,

Koristka²,

Keil³

et al. 2012

Antibodies

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Immunotherapy has emerged as an alternative strategy to treat malignancies in addition to conventional radio- and chemotherapy. There has been a plethora of evidence that the immune system is able to control tumor outgrowth and a number of strategies have been put forward to utilize this ability for immunotherapy. However, some of these strategies have not been very efficient and their success has been limited by tumor evasion mechanisms. A promising approach to engage effector cells of the immune system overcoming some of the escape mechanisms has been introduced more than two decades ago. This approach is based on bispecific antibodies. Here we summarize the evolution of bispecific antibodies, their improvement, remaining obstacles and some controversial reports.

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Engineering and characterization of a bispecific HER2 × EGFR‐binding affibody molecule

Abstract: HER2 (human epidermal-

Cited by 60 publications

References 46 publications

A Bispecific Enediyne-Energized Fusion Protein Containing Ligand-Based and Antibody-Based Oligopeptides against Epidermal Growth Factor Receptor and Human Epidermal Growth Factor Receptor 2 Shows Potent Antitumor Activity

A Bispecific Enediyne-Energized Fusion Protein Containing Ligand-Based and Antibody-Based Oligopeptides against Epidermal Growth Factor Receptor and Human Epidermal Growth Factor Receptor 2 Shows Potent Antitumor Activity

Affibody molecules: Engineered proteins for therapeutic, diagnostic and biotechnological applications

Cancer Immunotherapy by Retargeting of Immune Effector Cells via Recombinant Bispecific Antibody Constructs

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