@conference{
author = "Radomirović, Mirjana and Čolaković, Maša and Pismestrović, Marina and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković Veličković, Tanja",
year = "2022",
abstract = "Tropomyosin (TPM) is considered as a major allergen among different shellfish species.
Therefore, the development of methods for quantifying TPM in food products iscrucial for
allergic persons. Several extraction buffers were tested for their efficiency in recovering
proteins from fresh frozen and cooked razor mud shrimp during 2 and 24 hours of
extraction. The protein content was quantified using the Bradford protein assay. SDSPAGE
was used for protein profiling of soluble extracts. Sandwich ELISA was developed
and used to quantify TPM content. None of the extraction buffers showed a significant
difference in total protein content between 2 and 24 hours of extraction. Significantly
fewer proteins were extracted from cooked shrimp compared to the raw shrimp. ELISA
quantification showed that phosphate-buffered saline (PBS) containing 1 M NaCl (PBSN)
and carbonate buffer, pH 10, extracted approximately 6 times higher amount of
tropomyosin in comparison to PBS, highlighting the importance of choosing the
appropriate extraction buffer for the precise quantification of TPM. Traditionally used
extraction buffer PBS could significantly underestimate shrimp TPM content., Tropomiozin (TPM) se smatra glavnim alergenom morskih plodova. Razvoj metoda za kvantifikaciju TPM-a u prehrambenim proizvodima je ključan za alergične osobe. Nekoliko pufera za ekstrakciju je upoređeno u pogledu efikasnosti ekstrakcije protein iz sveže zamrznutih i kuvanih gambora, tokom 2 i 24 časa. Sadržaj proteina u solubilnim ekstraktima je određen Bredfordovom metodom. Proteinski profil ekstrakata je analiziran SDS-PAGE metodom. Za kvantifikaciju TPM-a je razvijena sendvič ELISA metoda. Nijedan puffer nije pokazao značajnu razliku u količini ekstrahovanih protein nakon 2 i 24 časa ekstrakcije. U poređenju sa sirovim, značajno manje proteina je ekstrahovano iz kuvanih gambora. Kvantifikacija TPM-a ELISA metodom je pokazala da se fosfatom puferisanim fiziološkim rastvorom (PBS) koji sadrži 1 M NaCl (PBSN) i karbonatnim
puferom, pH 10, ekstrahuje oko 6 puta više TPM-a u poređenju sa PBS-om. Ovo ukazuje
na značaj odabira adekvatnog pufera za ekstrakciju TPM-a, jer se upotrebom tradicionalno
korišćenih pufera može potceniti koncentracija TPM-a u gamborima.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "58th Meeting of the Serbian Chemical Society - Book of abstracts, Proceedings, June 9-10, 2022 Belgrade / 58. Savetovanje Srpskog hemijskog društva, Kratki izvodi radova, Knjiga radova, 9. i 10. jun 2022. godine, Beograd",
title = "Optimization of extraction conditions of tropomyosin from razor mud shrimp and its quantification by developed ELISA",
pages = "64-64",
url = "https://hdl.handle.net/21.15107/rcub_cer_6823"
}
