Optimizacija ekstrakcije proteinov za detekcijo z encimskoimunskim testom (ELISA) v modelnih vzorcih proteinskih barv

Penko, Ana (author), Kralj Cigić, Irena (mentor)

Namen tega dela je bila optimizacija zaznave jajčnega proteina ovalbumina s testom ELISA v modelnih vzorcih proteinskih barv dveh bakrovih pigmentov – malahita in pigmenta verdigris. Gre za prvo poglobljeno in sistematično raziskavo v dediščinski znanosti, v kateri smo s spremljanjem koncentracije bakrovih ionov iz pigmentov barv poskušali razumeti njihov vpliv na razgradnjo barv in kako razgradnja ter postopki priprave vzorcev (primerjava nečiščenih in čiščenih vzorcev) pri posameznih korakih testa ELISA vplivajo na končno zaznavo proteinskega materiala. Eden najpomembnejših doprinosov te študije k dediščinski znanosti je izboljšanje meje zaznave ovalbumina v barvi pigmenta verdigris s testom ELISA za 10-krat v razgrajenih in 100-krat v nerazgrajenih modelnih vzorcih. Razlika vpliva čiščenja raztopin na zaznavo ovalbumina s testom ELISA med bakrovima pigmentoma malahita in pigmenta verdigris najverjetneje izhaja iz razlik v topnosti teh pigmentov.

• Language: Slovenian
• Keywords: kulturna dediščina, proteini, ELISA, optimizacija, diski CIMmic
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2021
• PID: 20.500.12556/RUL-128215
• COBISS.SI-ID: 73945859
• Publication date in RUL: 06.07.2021

Secondary language

• Language: English
• Title: Optimisation of protein extraction for detection with enzyme-linked immunosorbent assay (ELISA) in protein paint model samples

Abstract

The aim of this work was optimisation of ovalbumin (egg protein) detection in protein paint model samples containing two different copper pigments – malachite and verdigris, using ELISA assay. This is first in-depth and systematic study in heritage science where the effect of copper ions on paint degradation was studied by ion concentration monitoring. We also studied how degradation and sample preparation procedures (comparison of non-purified and purified samples) effect on protein detection using ELISA. One of the most important contributions of this study to heritage science is 100-fold improvement of ovalbumin detection limit in non-degraded verdigris paint and 10-fold improvement in degraded verdigris paint. Difference between purifying effect on ovalbumin detection using ELISA in malachite and verdigris paint most likely originates from solubility difference of those pigments.

• Keywords: cultural heritage, proteins, ELISA, optimisation, CIMmic disks