The project aims at the innovative analytical approach based on biofunctionalized magnetic micro/nanoparticles combined with microfluidic analytical device (m-TAS). Benefits provided by both systems open the feasibility to use this microanalyzator for effective separation of bacterial proteins from highly complex mixture. The analyses will be focused on significant attributes of virulence and pathogenesis within intracellular parasite. New analytical strategy combined with mass spectrometry (MS) enables to isolate and characterize the structure of low-abundant glykoproteins, phosphoproteins and in other way modified proteins from cell lysate or membrane fractions of bacteria and identify biomolecules predicting the virulence of bacteria strains. The immobilized magnetic affinity reactor (IMAR) with specific ligand used for isolation and purification of proteins significantly facilitates the identification of protein fingerprint by MS even for bacterial protein in submicromolar concentration. Comparative analyses will be performed on the target proteins isolated from two bacterial strains of Francisella tularensis differing in virulence. (en)
Vyvinout a validovat mikroprůtokový analytický systém, jehož integrální součástí jsou specifickým ligandem modifikované magnetické mikro/nanočástice; tento systém použít k identifikaci a strukturní charakterizaci bakteriálních proteinů majících vztah k virulenci a patogenezi bakterií.