Targeting Human Thrombus by Liposomes Modified with Anti-Fibrin Protein Binders

0521340 - BTÚ 2020 RIV CH eng J - Journal Article
Petroková, Hana - Mašek, J. - Kuchař, Milan - Wunschova, A. V. - Štikarová, J. - Bartheldyová, E. - Kulich, P. - Hubatka, F. - Kotouček, J. - Knötigová, P.T. - Vohlidalova, E. - Hezova, R. - Mašková, E. - Macaulay, S. - Dyr, J. E. - Raška, M. - Mikulik, R. - Malý, Petr - Turánek, J.
Targeting Human Thrombus by Liposomes Modified with Anti-Fibrin Protein Binders.
Pharmaceutics. Roč. 11, č. 12 (2019), č. článku 642. E-ISSN 1999-4923
R&D Projects: GA MZd(CZ) NV16-30299A; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:86652036
Keywords : fibrin * thrombus imaging * binding protein * thrombus targeting
OECD category: Pharmacology and pharmacy
Impact factor: 4.421, year: 2019
Method of publishing: Open access
https://www.mdpi.com/1999-4923/11/12/642

Development of tools for direct thrombus imaging represents a key step for diagnosis and treatment of stroke. Nanoliposomal carriers of contrast agents and thrombolytics can be functionalized to target blood thrombi by small protein binders with selectivity for fibrin domains uniquely formed on insoluble fibrin. We employed a highly complex combinatorial library derived from scaffold of 46 amino acid albumin-binding domain (ABD) of streptococcal protein G, and ribosome display, to identify variants recognizing fibrin cloth in human thrombus. We constructed a recombinant target as a stretch of three identical fibrin fragments of 16 amino acid peptide of the B beta chain fused to TolA protein. Ribosome display selection followed by large-scale Enzyme-Linked ImmunoSorbent Assay (ELISA) screening provided four protein variants preferentially binding to insoluble form of human fibrin. The most specific binder variant D7 was further modified by C-terminal FLAG/His-Tag or double His-tag for the attachment onto the surface of nanoliposomes via metallochelating bond. D7-His-nanoliposomes were tested using in vitro flow model of coronary artery and their binding to fibrin fibers was demonstrated by confocal and electron microscopy. Thus, we present here the concept of fibrin-targeted binders as a platform for functionalization of nanoliposomes in the development of advanced imaging tools and future theranostics.
Permanent Link: http://hdl.handle.net/11104/0305969