Kromatografsko pročišćavanje biomolekula i njihova karakterizacija
|2013-11||8193||CHROBIO||15.09.2014 - 14.09.2017||Završio||676.340,51 Kn|
Interdisciplinarni, Prirodne znanosti, Biomedicina i zdravstvo, Biotehničke znanostiZnanstvena polja:
Sveučilište u ZagrebuSuradnici:
Jelena Ivančić Jelečki, Tihana Kurtović, Maja Markušić, Beata Halassy, DUBRAVKO FORČIĆ, Dora SvibenKljučne riječi:
chromatography, viruses, plasma proteins, snake venom, monoliths, bioanalytics, aggregationSažetak:
Aim of the proposed research is the design of non-invasive chromatographic methods suitable for purification and concentration of biomolecules (viruses and proteins) and their characterization. Purification and analysis methods will be focused on measles virus, snake venom and antivenom proteins, and human plasma proteins. Measles virus is used as a vaccine and is found very suitable as a gene vector and as an oncolytic virotherapeutic, with the last two applications requiring large amounts of the virus. Venom is an antigen for antivenom production and both require careful analysis in order to obtain the final product of high quality. Proteins from plasma, namely albumin and immunoglobulin, are of great medical importance and their purification and stability are of utmost significance. All these biomolecules are important as biotechnological products and are subjected to rigorous quality requirements to ensure their safe usage. Therefore, non-invasive purification methods upgrading their quality and sustainable production and ensuring stability of these fragile biomolecules are essential. This project will base purification on chromatography using monolith stationary phases which are featured by their suitability for biomolecules due to high porosity, high binding capacity and convective based transport. Ion-exchange and affinity chromatography will be employed and various conditions tested for obtaining target material with high purity and yield. Also, various formulation conditions enabling preservation of virus viability and protein functionality and stability throughout the process will be tested. The analysis of the virus infectivity and protein functionality, removal of contaminants and the overall procedure effectiveness will be done by classical biochemical and virological methods as well as by some novel methods such as nanoparticle analysis that enables detection of virus and protein aggregates (NanoSight).