magistrsko delo
Ana Grobler (Author), Aleš Podgornik (Reviewer), Romana Marinšek-Logar (Mentor), Luka Jeromel (Co-mentor)

Abstract

Biološke učinkovine zahtevajo izjemno čistost, vendar je postopek čiščenja zelo zahteven. Da bi dosegli ustrezne ravni čistosti je treba izvesti več kromatografskih korakov. Tekočinska kromatografija je še vedno glavna tehnika izbire, ki se uporablja za čiščenje bioloških snovi. Kakovost spakirane kromatografske kolone ima pomembno vlogo pri izvedbi kromatografskega koraka. V tem študiji smo določili vpliv pakiranja kromatografskih kolon na kakovost produkta. V ta namen smo spakirali deset kromatografskih kolon, ki so se razlikovale v kakovosti. Spakirane kromatografske kolone smo testirali pred in po vsaki separaciji s pomočjo pulznega testa, da bi ocenili njihovo kakovost, opredeljeno s tremi parametri: število teoretičnih podov, višina teoretičnega poda in asimetrija. Namerno smo spakirali kolone, ki so se razlikovale v vseh parametrih in odstopale od priporočenih vrednosti. Po pripravi kolon smo na vseh izvedli separacijo z uporabo hidrofobne interakcijske kromatografij. Analiza eluatov je pokazala, da je vsebnost nečistoč skladna med posameznimi kromatografskimi kolonami. Ob upoštevanju negotovosti analize smo zaključili, da so vse spakirane kromatografske kolone enakovredne glede čiščenja proteina, poleg tega je vsaka od kromatografskih kolon protein očistila do ustrezne ravni čistosti.

Keywords

preparativna kromatografija;hidrofobna interakcijska kromatografija;kromatografske kolone;pakiranje kromatografskih kolon;test kolone;parametri kvalitete kolone;separacija;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [A. Grobler]
UDC: 543.544.5:66.01
COBISS: 4889464 Link will open in a new window
Views: 1138
Downloads: 293
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Other data

Secondary language: English
Secondary title: Impact of chromatographic column packing on product quality
Secondary abstract: Biopharmaceuticals require exceptional purity, however, purification process is a challenging task. To achieve the required levels of purity, several chromatographic steps need to be implemented. Liquid chromatography is still main technique of choice used for purification of biological substances. Quality of packed chromatographic column plays a significant role in the performance of the chromatographic step. In the present study we determined the impact of packing of chromatographic columns on product quality. For this purpose, we packed ten columns differently in order to obtain various quality of packed chromatographic columns. After the columns were packed, we tested them before and after each separation using a pulse test in order to assess their quality defined by three measures: number of plates, height equivalent to the theoretical plate and asymmetry. We intentionally packed columns that differed in all parameters and deviated from recommended values. After column preparation, a separation using the hydrophobic interaction chromatography was carried out on each of packed columns. The analysis of the eluates showed that the content of impurities was consistent between individual chromatographic columns. Taking into account the uncertainty of the analysis, we concluded, that all of the packed columns are equivalent concerning the purification of product; moreover, each of the columns purified the product to proper levels of purity.
Secondary keywords: preparative chromatography;hydrophobic interaction chromatography;chromatographic column;packing chromatographic column;column test;column quality parameters;separations;
Type (COBISS): Master's thesis/paper
Study programme: 0
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Študij mikrobiologije
Pages: XIV, 84 f., [10] f. pril.
ID: 10918717