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Targeting Human Thrombus by Liposomes Modified with Anti-Fibrin Protein Binders
- Source :
- Pharmaceutics, Volume 11, Issue 12
- Publication Year :
- 2019
- Publisher :
- MDPI AG, 2019.
-
Abstract
- 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<br />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.
- Subjects :
- Pharmaceutical Science
binding protein
02 engineering and technology
ABD scaffold
Article
Fibrin
law.invention
03 medical and health sciences
law
medicine
combinatorial library
fibrin
thrombus imaging
Thrombus
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
Liposome
metallochelation
biology
Binding protein
021001 nanoscience & nanotechnology
medicine.disease
In vitro
Amino acid
chemistry
liposome
Ribosome display
biology.protein
Recombinant DNA
Biophysics
thrombus targeting
fibrinogen Bβ chain
0210 nano-technology
Subjects
Details
- ISSN :
- 19994923
- Volume :
- 11
- Database :
- OpenAIRE
- Journal :
- Pharmaceutics
- Accession number :
- edsair.doi.dedup.....363f7970b53a778f9c5c698ffcdf17fe