magistrsko delo
Urška Poglajen (Author), Aleš Podgornik (Mentor), Aleš Podgornik (Thesis defence commission member), Mojca Narat (Thesis defence commission member), Iztok Prislan (Thesis defence commission member)

Abstract

Monoklonska protitelesa so veja bioloških zdravil, ki so proizvedena v različnih celičnih sistemih s pomočjo tehnologije rekombinantne DNA. Protitelesa so glikoproteini, ki so zelo pomembni pri protitelesnem imunskem odzivu. Med proizvodnjo velikih in kompleksnih proteinov so nujno potrebne post-translacijske modifikacije (PTM), za njihovo biološko aktivnost. Med najbolj pomembne PTM spada glikozilacija, kjer poteka pripenjanje sladkorjev na aminokislinske ostanke protitelesa. Glikozilacija je zelo pomembna modifikacija terapevtskih proteinov, s katero lahko spreminjamo izkoristek, učinkovitost, topnost, stabilnost (proteoliza), imunogenost in hitrost izločanja učinkovine iz telesa. Prav zato je temeljita karakterizacija vsebnosti sladkornih ostankov, strukture sladkorne verige in glikozilacijskih mest, ki so prisotna na protitelesu, ključnega pomena za zagotavljanje varnosti in učinkovitosti zdravila. Zaradi kompleksnosti in zamudnosti dosedanjih analitskih metod je smiselno poiskati alternativne tehnike. Ker so glikanske strukture v notranjosti protiteles, jih je potrebno predobdelati, kar lahko naredimo na različne načine. Poleg encimske metode, kjer pride do cepitve glikanov, je možna tudi termična predobdelava, pri kateri pride do delnega odprtja strukture. Primarni cilj eksperimentalnega dela je bil poiskati pogoje termične predobdelave, pri katerih ne bi prišlo do agregacije. Prav tako smo želeli poiskati metodo, s katero bi potrdili odprtje strukture molekule in lokalizirali mesto, na katerem pride do razprtja.

Keywords

monoklonska protitelesa;glikozilacija;termična obremenitev mAb;ločevanje izooblik;lektini;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [U. Poglajen]
UDC: 602.44:543.544.14/15:615.3(043.2)
COBISS: 9215865 Link will open in a new window
Views: 766
Downloads: 202
Average score: 0 (0 votes)
Metadata: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Other data

Secondary language: English
Secondary title: ǂThe ǂimpact of the pre-treatment of monoclonal antibodies on separation of isoforms with ion exchange chromatography
Secondary abstract: Monoclonal antibodies are a branch of biological drugs that are produced in different cellular systems using recombinant DNA technology. Antibodies are glycoproteins and they are very important in the antibody immune response. During the production of large and complex proteins, post-translational modifications (PTM) are essential for their biological activity. Among the most important PTMs are glycosylation, where the attachment of sugars to the amino acid residues of the antibody takes place. Glycosylation is a very critical modification of therapeutic proteins known to change efficacy, solubility, stability (especially prior to proteolysis), immunogenicity and the rate of clearance of the active substance from the body. For this reason, the thorough characterization of the sugar residues content, the sugar chain structure and glycosylation sites that are present in the antibody is essential to ensure the safety and efficacy of the drug. Due to the complexity and obscurity of previous analytical methods, it is useful to find alternative techniques. Because glycan structures are inside the antibodies, they need to be pre-treated, which can be done in different ways. In addition to the enzymatic method where the glycans are split, thermal pretreatment is also possible, in which partial opening of the structure occurs. The primary goal of the experimental work was to find the conditions of thermal pretreatment in which there would be no aggregation. Likewise, we wanted to find a method that would confirm the opening of the molecular structure and the position where the dispersion occurs.
Secondary keywords: monoclonal antibodies;glycosylation;thermal pretreatment of mAb;separation of isoforms;lectins;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije
Pages: XII, 56 f., [6] f. pril.
ID: 11141142