Biotehnološka proizvodnja terapijskog proteina filgrastima

undergraduate thesis

• Title (english): Biotechnological production of theraphy protein filgrastim
• Author: Antonija Vrbić
• Mentor: Jagoda Šušković (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Antibiotic, Enzyme, Probiotic and Starter Cultures Technology); Zagreb
• Defense date and country: 2011-06-29, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Filgrastim je rekombinantni terapijski protein koji djeluje kao faktor stimulacije humanih granulocitnih kolonija. Prvenstveno se koristi kako bi se smanjila učestalost i trajanje kronične neutropenije i pridruženih komplikacija. Razvijen je u Hospiri kao generik Amgen filgrastima. U ovom radu opisana je proizvodnja filgrastima dizajniranog prema humanom proteinu G-CSF i njegove karakteristike. Za proizvodnju se koriste tehnike rekombinante DNK tehnologije te stanica Escherichia coli kako bi se dobio protein što sličniji humanom i sačuvala njegova svojstva. Filgrastim se proizvodi dalje submerznim uzgojem E.coli u više faza pri optimalnim fiziološkim uvjetima. Proces proizvodnje je podijeljen u četiri faze, koje čine uzgoj cjepiva I u Erlenmeyer tikvicama, zatim uzgoj cjepiva II za biosintezu u biorektoru manjeg volumena, biosinteza u bioreaktoru volumena 30 l te faze separacije i pročišćavanja. Budući da se proizvod nalazi u obliku inkluzijskih tijela, potrebne su višestruke separacije razmuljivanjem i centrifugiranjem, prvo same dobivene biomase zatim inkluzijskih tijela. Pri proizvodnji veliku važnost ima i kvaliteta vode koja je višestruko pročišćavana te se stoga uzorkovala voda za injekcije koja se koristi i provedena je procjena kvalitete vode.

Abstract (english)

Filgrastim is a recombinant therapeutic protein that acts as a human granulocyte- colony stimulating factor. It is primarily used to reduce the incidence and duration of severe neutropenia and associated complications. It is developed in Hospira, as a biosimilar according to Amgen Filgrastim. This paper describes the production of Filgrastim, designed according to human protein G-CSF, and its characteristics. Recombinant DNA technology techniques and the Escherichia coli cells are used for the production to obtain a protein similar to the human and preserve its properties. Filgrastim is produced by submerged cultivation of E.coli in several stages, under optimal physiological conditions. The production process is divided into four phases, which are growing vaccine I in Erlenmeyer flasks, then growing the vaccine II in the biosynthesis bioreactor of smaller volume, biosynthesis in bioreactor with the volume of 30 l, followed by separation and purification. Since the product is in the form of inclusion bodies, multiple separations by centrifugation are required, first of obtained biomass then inclusion bodies. In the production, the quality of water which is multiply purified is of great importance. Therefore, the water for injections was sampled and assessment of water quality was carried out.

• Keywords: biosinteza; G-CSF; filgrastim; Escherichia coli; rekombinantna DNK tehnologija
• Keywords (english): biosynthesis; G-CSF; Filgrastim; Escherichia coli; recombinant DNA technology
• Language: croatian
• URN:NBN: urn:nbn:hr:159:860472
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-19 11:52:16