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Title:Primerjava različnih vrst stekla za in situ karakterizacijo bioloških zdravil : diplomsko delo univerzitetnega študijskega programa I. stopnje
Authors:ID Jagodič, Maša (Author)
ID Zalar, Matja (Mentor) More about this mentor... New window
ID Bren, Urban (Comentor)
Files:.pdf UN_Jagodic_Masa_2025.pdf (4,28 MB)
MD5: 85360A650DE8098CEAF8C6688A43B899
 
Language:Slovenian
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:Biološka zdravila na osnovi proteinov se soočajo s problematiko stabilnosti pri različnih pogojih shranjevanja. Proteini so podvrženi agregaciji in razvijanju sekundarne strukture, ki povzročijo izgubo njihove primarne funkcije in lahko vodi do ogrožanja zdravja pacientov. Ti procesi so pospešeni pod stresnimi pogoji, kot sta povišana temperatura in , izpostavljenosti svetlobi. Trenutne metode za ocenjevanje strukturne integritete proteinov namreč zahtevajo predpripravo vzorcev in redčenje, pri čemer je povečana možnost človeške napake. Mednje spadajo spektroskopske metode kot so krožni dihroizem (CD), ultravijolična-vidna spektroskopija (UV-Vis) in Fourierjeva transformacijska infrardeča spektroskopija (FTIR). Zaradi inertnosti, so biološka zdravila najpogosteje shranjena v steklenih vialah, ki omogočajo izvajanje spektroskopskih metod. Naš cilj je bil preveriti, katere vrste stekla omogočajo razvoj metode na podlagi CD spektroskopije, s katero bo omogočena in situ analiza proteinov. Primerjali smo vpliv različnih vrst stekla na razvijanje sekundarne strukture proteina.V diplomskem delu smo preučevali tudi vpliv robnih pogojev na strukturo lizocima. To smo opazovali s snemanjem v kvarčnih kivetah, HPLC in farmacevtskih vialah. Ugotovili smo, da pri spremenjenih pogojih, ne pride do razvijanja sekundarne strukture lizocima, ko pa se struktura spremeni, takoj nastopi agregacija. Primerjava stekla pripelje do zaključka, da je za snemanje v CD spektrometru pričakovano najbolj ugodna kvarčna kiveta. HPLC viala zaradi samo absorpcije in napetosti nad mejno vrednostjo nasičenja detektorja, ne omogoča izvajanje meritev. Tako kvarčna kiveta, ki je specializirana za merjenje v CD spektru, kot tudi farmacevtska viala zadovoljita pogoje za snemanje v CD spektrometru.
Keywords:biološka zdravila, krožni dihroizem(CD), in situ karakterizacija, shranjevanje bioloških zdravil, vrste stekla
Place of publishing:Maribor
Place of performance:Maribor
Publisher:[M. Jagodič]
Year of publishing:2025
Number of pages:1 spletni vir (1 datoteka PDF (X, 33 str.))
PID:20.500.12556/DKUM-94289 New window
UDC:547.96.032.1:615.014.4(043.2)
COBISS.SI-ID:249588227 New window
Publication date in DKUM:09.09.2025
Views:132
Downloads:37
Metadata:XML DC-XML DC-RDF
Categories:KTFMB - FKKT
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:12.08.2025

Secondary language

Language:English
Title:Comparison of different types of glass for in situ characterization of biological drugs
Abstract:Protein-based biologics face stability issues under various storage conditions. Proteins are prone to aggregation and the development of secondary structure, which impair their primary function and pose a risk to patient health. These processes are accelerated by elevated temperatures, exposure to light, etc. Current methods for assessing protein structural integrity require sample preparation and dilution, which increases the potential for human error. These include spectroscopic methods such as circular dichroism (CD), ultraviolet-visible spectroscopy (UV-Vis), and Fourier transform infrared spectroscopy (FTIR). Due to their inertness, biologics are most often stored in glass vials, which enable the use of spectroscopic methods. Our goal was to determine which types of glass enable the development of a CD spectroscopy-based method for in situ protein analysis. We compared the effects of different glass types on CD spectroscopy.We compared quality of spectra obtained in quartz cuvettes, pharmaceutical grade vials and HPLC vials. In this thesis we also studied the influence of stress conditions on the structure of lysozyme. We found that under altered conditions, lysozyme does not develop a secondary structure, and when a change in structure does occur, it leads to aggregation. Comparison of the spectra quality, indicated thatthe best results are obtained in quartz cuvettes, as one would expect. Nevertheless, the tested pharmaceutical grade vials performed well, allowing for in-situ analysis of tertiary and secondary structure. The HPLC vial, however, does not allow measurements of CD spectra due to absorption of UV spectra exceeding the saturation limit of the detector.
Keywords:biological drugs, circular dichroism (CD), in situ characterization, biological drug storage, glass type


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