Optimization of extraction conditions of tropomyosin from razor mud shrimp and its quantification by developed ELISA
Аутори
Radomirović, Mirjana Ž.Čolaković, Maša
Pismestrović, Marina
Gligorijević, Nikola
Stanić-Vučinić, Dragana
Rajković, Andreja
Ćirković-Veličković, Tanja
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Tropomyosin (TPM) is considered a major allergen among different shellfish species. Therefore, the development of methods for quantifying TPM in food products is crucial 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. SDS-PAGE 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.
Извор:
58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings, 2022, 64-64Издавач:
- Beograd : Srpsko hemijsko društvo
Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200168 (Универзитет у Београду, Хемијски факултет) (RS-MESTD-inst-2020-200168)
- ShellPCR - Development of Elisa and Immuno-PCR for Sensitive and Specific Detection of Shellfish Tropomyosin (RS-ScienceFundRS-Dijaspora-6504499)
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - CONF AU - Radomirović, Mirjana Ž. AU - Čolaković, Maša AU - Pismestrović, Marina AU - Gligorijević, Nikola AU - Stanić-Vučinić, Dragana AU - Rajković, Andreja AU - Ćirković-Veličković, Tanja PY - 2022 UR - http://cherry.chem.bg.ac.rs/handle/123456789/6027 AB - Tropomyosin (TPM) is considered a major allergen among different shellfish species. Therefore, the development of methods for quantifying TPM in food products is crucial 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. SDS-PAGE 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. PB - Beograd : Srpsko hemijsko društvo C3 - 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings T1 - Optimization of extraction conditions of tropomyosin from razor mud shrimp and its quantification by developed ELISA SP - 64 EP - 64 UR - https://hdl.handle.net/21.15107/rcub_cherry_6027 ER -
@conference{ author = "Radomirović, Mirjana Ž. 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 a major allergen among different shellfish species. Therefore, the development of methods for quantifying TPM in food products is crucial 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. SDS-PAGE 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.", publisher = "Beograd : Srpsko hemijsko društvo", journal = "58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings", 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_cherry_6027" }
Radomirović, M. Ž., Čolaković, M., Pismestrović, M., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2022). Optimization of extraction conditions of tropomyosin from razor mud shrimp and its quantification by developed ELISA. in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings Beograd : Srpsko hemijsko društvo., 64-64. https://hdl.handle.net/21.15107/rcub_cherry_6027
Radomirović MŽ, Čolaković M, Pismestrović M, Gligorijević N, Stanić-Vučinić D, Rajković A, Ćirković-Veličković T. Optimization of extraction conditions of tropomyosin from razor mud shrimp and its quantification by developed ELISA. in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings. 2022;:64-64. https://hdl.handle.net/21.15107/rcub_cherry_6027 .
Radomirović, Mirjana Ž., Čolaković, Maša, Pismestrović, Marina, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković-Veličković, Tanja, "Optimization of extraction conditions of tropomyosin from razor mud shrimp and its quantification by developed ELISA" in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings (2022):64-64, https://hdl.handle.net/21.15107/rcub_cherry_6027 .