Josef Masek
Masaryk University, Czech Republic
Title: Protein binder-decorated liposomes targeted to fi brin for direct thrombus imaging
Biography
Biography: Josef Masek
Abstract
Direct thrombus imaging represents an advanced approach for stroke diagnosis. Liposomes are an almost ideal and clinically proven platform for the construction of both, diagnostic contrast agents and nanotherapeutics. We constructed thrombus-targeted liposomal systems using protein binders with selectivity for fibrin domains uniquely formed on insoluble fibrin. Protein binders were selected using a highly complex combinatorial library derived from the scaffold of 46 amino acid albumin-binding domain of protein G, and ribosome display. D7 variant of protein binder was modified by His-Tag or double His-Tag enabling surface decoration of liposomes via metallochelating bond. The surface modification of liposomes was confirmed using different techniques including transmission electron microscopy, immunogold staining, dynamic light scattering, and isothermal titration. D7-modified liposomes were shown to bind to fibrin fibrils of the thrombus using laser scanning confocal microscopy and electron microscopy. Static conditions, as well as in-vitro flow model of middle cerebral artery, were tested. In conclusion, we present here the concept of liposomes functionalized with fibrin-targeted binders for direct thrombus imaging.