DARPins as bispecific receptor antagonists analyzed for immunoglobulin E receptor blockage

Eggel, Alexander; Baumann, Michael J; Amstutz, Patrick; Stadler, Beda M; Vogel, Monique (2009). DARPins as bispecific receptor antagonists analyzed for immunoglobulin E receptor blockage. Journal of molecular biology, 393(3), pp. 598-607. Amsterdam: Elsevier 10.1016/j.jmb.2009.08.014

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The concept of multispecific antibodies is of high therapeutic interest but has failed to produce pharmaceutical products due to the poor biophysical properties of such molecules. Here, we propose an alternative and simple way to generate bispecific binding molecules using designed ankyrin repeat proteins (DARPins). For this purpose, monovalent DARPins with different epitope specificities were selected against the alpha chain of the high-affinity receptor for human immunoglobulin E (IgE) (FcepsilonRIalpha). Two of the isolated binders interfering with IgE binding to the receptor were joined to each other or to themselves via a flexible protein linker. The resulting bivalent and bispecific DARPins were tested for their ability to prevent allergen-induced cell degranulation using rat basophilic leukemia cells stably transfected with human FcepsilonRIalpha. The bispecific DARPin construct was the most potent one, efficiently blocking the IgE-FcepsilonRI interaction and preventing the release of proinflammatory mediators. Noteworthy, the multivalent and multispecific DARPin construct did not show any alteration of the beneficial biophysical properties of the monovalent parental DARPins. Hence, bispecific DARPins may be used to generate receptor antagonists simultaneously targeting different epitopes on the same molecule. Moreover, they easily overcome the limiting immunoglobulin binding paradigm (one binding molecule=one epitope) and thereby represent an alternative to monoclonal antibodies in cases where the immunoglobulin scaffold is unsuitable.

Item Type:	Journal Article (Original Article)
Division/Institute:	04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Institute for Immunology [discontinued]
UniBE Contributor:	Stadler, Beda Martin, Vogel, Monique
ISSN:	0022-2836
Publisher:	Elsevier
Language:	English
Submitter:	Factscience Import
Date Deposited:	04 Oct 2013 15:13
Last Modified:	05 Dec 2022 14:22
Publisher DOI:	10.1016/j.jmb.2009.08.014
PubMed ID:	19683003
Web of Science ID:	000271167400004
URI:	https://boris.unibe.ch/id/eprint/32344 (FactScience: 197445)