magistrsko delo
David Mlakar (Avtor), Aleš Podgornik (Mentor)

Povzetek

V današnjem času so protitelesa in fragmenti protiteles eni izmed najbolj pogostih in zaželenih biotehnoloških produktov, pri čemer je Pichia pastoris pogosto izbrana kot produkcijski organizem, saj gre za enega bolj preučenih mikroorganizmov z izredno dobrimi lastnostmi za industrijsko produkcijo. Pri načrtovanju bioprocesne sheme se pogosto osredotočamo na podoben vzorec korakov, vendar pa je za vsak proces potrebno razviti celovit, individualen pristop. Pri našem delu smo se zato posvetili individualnemu pristopu optimizacije gojenja kvasovke vrste Pichia pastoris na majhni bioreaktorski skali z namenom produkcije protitelesnih fragmentov Fcab. S spreminjanjem priprave inokuluma in načina inokulacije smo postopek standardizirali, s spreminjanjem pogojev gojenja, kot je stopnja redčenja, pa smo uspešno s kontinuirnim postopkom gojili kulturo, ki je ostala produkcijsko aktivna. Slednje smo dokazali z analizami izolatov produkta z metodo SDS-PAGE. Reverzibilna vezava fragmenta Fcab s proteinom A nam je omogočila uporabo afinitetne kromatografske metode tako za namen njihove izolacije, hkrati pa smo z rezultati lahko ocenili tudi končno koncentracijo produkta v bioreaktorju.

Ključne besede

kemostat;Pichia pastoris;fragment protiteles Fcab;afinitetna kromatografija,;SDS-PAGE;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.09 - Magistrsko delo
Organizacija: UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik: [D. Mlakar]
UDK: 606:616-097.3:602.4:602.3:582.282.23(043.2)
COBISS: 95344899 Povezava se bo odprla v novem oknu
Št. ogledov: 183
Št. prenosov: 38
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Production of antibody fragments in yeast Pichia pastoris in a continuous bioprocess
Sekundarni povzetek: Antibodies and their fragments are considered as one of the most desirable biotechnological products. Pichia pastoris is one of the more studied microorganisms, with extremely good properties for industrial production, which is the reason why it is often selected as the production organism of choice for such complex products. When constructing a bioprocess scheme, we often follow the same rules, but still have to take an individual approach with each one, as no organism or product responses the same way to an adjustment. In our work we took this individual approach of optimization for our small-scale cultivation of Pichia pastoris, that produced Fcab antibody fragments. With changes to the inoculum preparation and inoculation process, we were able to standardize the start of the process. Adjustments of the parameters, such as the dilution rate, enabled the continuous growth of Pichia pastoris which maintained its productivity. This was proven with the isolation of Fcab from the fermentation broth at the end of several successive small-scale bioprocesses and their successful detection with SDS-PAGE. Additionally, to the isolation, we also used the resulting data of HPLC detectors, to calculate the estimated concentrations of Fcab fragments in each vessel at the end of each successful process.
Sekundarne ključne besede: chemostat;Fcab antibody fragment;afinity chromatography;
Vrsta dela (COBISS): Magistrsko delo/naloga
Študijski program: 0
Konec prepovedi (OpenAIRE): 1970-01-01
Komentar na gradivo: Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije
Strani: X, 55 f., [1] f. pril.
ID: 14324702