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Optimizacija metode kapilarnega izoelektričnega fokusiranja za analizo treh novih bioloških molekul = Optimization of a capillary isoelectric focusing method for analysis of three new biological molecules : magistrski študijski program Industrijska farmacijaMarinček, EvaBiološka zdravila so večinoma proteinske molekule, ki imajo v primerjavi z majhnimi sinteznimi učinkovinami mnogo večjo molekulsko maso ter kompleksno zgradbo, zato sta proces proizvodnje in ... karakterizacija zgradbe v njihovem razvoju ključnega pomena. Terapevtska monoklonska protitelesa postajajo ena izmed najhitreje rastočih skupin bioloških zdravil. Monoklonska protitelesa so heterogene molekule po velikosti in naboju. Nabite različice monoklonskih protiteles so eden izmed kritičnih atributov kakovosti, ki jih je treba karakterizirati in količinsko opredeliti ter določiti njihov vpliv na varnost in učinkovitost zdravila. Kapilarno izoelektrično fokusiranje (cIEF) je ena izmed metod, ki se uporablja za analizo in karakterizacijo nabitih različic. Na podlagi izoelektrične točke se različice ločijo v glavne, kisle in bazične skupine in jih na elektroferogramu vidimo kot ločene vrhove. V magistrski nalogi smo proučili primernost generične metode izoelektričnega fokusiranja pri razvoju treh bioloških molekul. Ker pri eni molekuli nismo dosegli zadovoljive ločbe, smo metodo optimizirali tako, da smo spreminjali sestavo reakcijske mešanice in pogoje fokusiranja. V generični metodi uporabljamo farmalit, ki pokriva široko pH območje (3-10), koncentracijo sečnine 2,0 M in čas fokusiranja 6 minut. V optimizirani metodi smo ločljivost uspeli izboljšati z dodatkom farmalita, ki pokriva ožje pH območje (8-10,5) ter s podaljšanjem časa fokusiranja na 10 minut. Za vse tri molekule smo določili območje, v katerem je metoda linearna, in testirali vpliv razsoljevanja. Ugotovili smo tudi, da je metoda primerna za zaznavanje sprememb v nabitih različicah proteina, ko tega izpostavimo stresnim pogojem.Type of material - master's thesis ; adult, seriousPublication and manufacture - Ljubljana : [E. Marinček], 2022Language - slovenianCOBISS.SI-ID - 113641219
Author
Marinček, Eva
Other authors
Kerč, Janez |
Kocjan, Boštjan
Topics
Biološka zdravila |
monoklonska protitelesa |
kapilarna elektroforeza |
izoelektrično fokusiranje |
nabite različice
Library/institution |
City | Acronym | For loan | Other holdings |
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Faculty of Pharmacy, Ljubljana | Ljubljana | FFALJ |
reading room 1 cop.
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Marinček, Eva | |
Kerč, Janez | 12942 |
Kocjan, Boštjan | 23430 |
Source: Personal bibliographies
and: SICRIS
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