20.500.12556/RUL-148488 Primerjava naprav za določanje koncentracije in viabilnosti celic CHO v pripravljalni fazi bioprocesa Comparison of different devices for determining cell concentration and viability in the upstream bioprocess using CHO cells. Industrijski bioproces gojenja celic CHO zahteva natančno določanje koncentracije celic in njihove viabilnosti zaradi številnih pomembnih razlogov. Naprava Vi-Cell XR, ki se trenutno uporablja v razvojnem oddelku bioloških učinkovin Lek d.d. Biofarmacevtike Mengeš, je bila umaknjena iz prodaje, kar predstavlja težavo zaradi pomanjkanja vzdrževanja in programske podpore. Iz tega razloga je bila izvedena primerjava dveh potencialnih nadomestnih naprav - Norma XS, ki temelji na metodi difrakcije svetlobe, in Vi-Cell BLU, ki temelji na barvanju s tripanskim modrilom. Na podlagi preizkusov in analize rezultatov je bilo ugotovljeno, da naprava Vi-Cell BLU ustrezno izpolnjuje vse testirane validacijske parametre za štetje celic in določanje njihove viabilnosti. Ta naprava je tudi natančnejša in bolj točna od dosedanje naprave Vi-Cell XR. Metoda barvanja s tripanskim modrilom se je izkazala kot primernejša od metode difrakcije svetlobe. Naprava Norma XS se kljub izjemni natančnosti ni izkazala za primerno. Kljub temu ima naprava potencial zaradi izjemno nizkih volumnov vzorcev in visoke natančnosti. Naprava, ki bo nadomestila Vi-Cell XR, mora biti vsaj enako točna in natančna ter omogočati boljše analize - vsem tem kriterijem zadostuje naprava Vi-Cell BLU. The industrial bioprocess of CHO cell cultivation requires accurate determination of cell concentration and viability for various important reasons. The Vi-Cell XR device, currently used in the Department of biological products development at Lek d.d. Biofarmacevtika Mengeš, has been discontinued, posing a problem due to the lack of maintenance and software support. Therefore, a comparison of two potential alternative devices was conducted: Norma XS, based on light diffraction method, and Vi-Cell BLU, based on trypan blue staining. Based on tests and analysis of results, it was found that the Vi-Cell BLU device adequately meets all tested validation parameters for cell counting and viability determination. This device is also more precise and accurate than the previous Vi-Cell XR device. The trypan blue staining method proved to be more suitable than the light diffraction method. Despite its exceptional precision, the Norma XS device was not deemed suitable due to imprecise measurements. Nevertheless, it has potential due to its low sample volumes and high precision if corrections were applied. A device that will replace the Vi-Cell XR must be at least equally accurate and precise, enabling improved analyses, process optimization, and informed decision-making, and the Vi-Cell BLU device meets all of these criteria. Industrijski bioproces celice CHO koncentracija in viabilnost celic Industrial bioprocess CHO cells cell culture cell concentration and viability true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2023-08-25 07:15:35 2023-08-25 07:15:38 2023-09-15 12:12:58 0000-00-00 00:00:00 2023 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 227515 162385411 34172.pdf 34172.pdf 1 408E0F7B61E13A22B518ECF5BD7EAFAE ff63d016952cecea1dce7375664887ab355fff3bf594ff42e2cff5112057a68d 5e158d50-4306-11ee-b9d7-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=172739 Biotehniška fakulteta 0 0 0