RESUMEN
We optimized a method for the preparation and purification of self-assembled protein nanocontainers EPN, the latest achievement in protein engineering. These nanocontainers are highly stable and provide the possibility of highly specific loading of a protein therapeutic drug. The described technique can be proposed as a tool for production of nanocontainers in a prokaryotic system. The obtained nanocontainers for the delivery of protein preparations can be used in the treatment of chronic, autoimmune, and oncological diseases.
Asunto(s)
Proteínas Recombinantes/biosíntesis , Nanoestructuras/química , Ingeniería de Proteínas/métodosRESUMEN
Previous data showed that myelin-reactive autoantibodies found in patients with multiple sclerosis and mice with experimental autoimmune encephalomyelitis recognize and hydrolyze various fragments of myelin basic protein (MBP). Moreover, antibody-mediated cleavage of the encephalithogenic fragment MBP81-103 flanked with two fluorescent proteins can serve as a new biomarker of multiple sclerosis. Here we describe creation of the next generation of this biomarker based on antibody-dependent degradation of a new chemically synthesized fluorescent substrate with resonance energy transfer that contains fluorophore Cy5 and quencher QXL680 separated by MBP81-99 protein (Cy5-MBP81-99-QXL680). This substrate is degraded during incubation with purified antibodies and B cells from patients with multiple sclerosis, but not healthy volunteers.
