Tropomyosin quantification in seafood samples-right choice of standard makes a difference
Аутори
Krstić-Ristivojević, Maja![](/themes/MirageCherry/images/orcid.png)
Jovanović, Vesna B.
![](/themes/MirageCherry/images/orcid.png)
Radomirović, Mirjana Ž.
![](/themes/MirageCherry/images/orcid.png)
Trifunović, Olga
Stanić-Vučinić, Dragana
![](/themes/MirageCherry/images/orcid.png)
Ćirković Veličković, Tanja
![](/themes/MirageCherry/images/orcid.png)
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
In the last 50 years, the annual per capita consumption of seafood products worldwide has more than doubled, from almost 10 kg in 1960 to over 20 kg in 2014. Seafood protein is an essential part of the diet in many countries, particularly where total protein intake is low [1]. However, as defined by the European Community, fish, and shellfish tropomyosins (TPM) are major allergens and major causes of anaphylaxis [2]. The increasing prevalence of food allergies is consistent with the increasing pollution of soil and water with plastic particles. To investigate the potential link between increasing plastic pollution and increasing food allergy prevalence, we aim to develop methods for precise and accurate monitoring of allergens and plastic in real seafood samples.
TPM was isolated from shrimp (Litopenaeus vannamei), clams (Venerupis philippinarum), and mussels (Mytilus galloprovincialis). The obtained in-house TPM proteins from three different sources were resolved using two-dimensiona...l polyacrylamide gel electrophoresis (2D-PAGE). The concentration of TPM in seashell samples from different geographical origin was determined using a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with prior optimization of adequate TPM standard curve using commercial and non-commercial in-house prepared TPM standards.
TPM standards resolved via 2D-PAGE revealed the presence of two isoforms of shrimp and mussels TPM standard, one dominant and one less abundant isoform. Two isoforms from both seafood sources, shrimp and mussels, are slightly different in molecular weight and pI value. As for the TPM standard obtained from clams, the 2D electrophoregram showed possibly eight isoforms with small differences in mass and pI values. Furthermore, the presence of three dominant isoforms can be observed that differ slightly in molecular mass, while other isoforms also differ in pI value. The ELISA results, regarding TPM standard curve optimization, showed that in both the commercial shrimp TPM and in-house shrimp TPM standards, sigmoidal concentration dependence is present in a range of 50 to 0.05 ng/ml, using serial double dilutions. On the other hand, TPM standards isolated from mussels and clams show sigmoidal concentration dependence in the range of 45 to 0.044 μg/ml with using the identical combination of capture and detection antibodies and serial double dilutions. TPM concentrations in clams and mussel samples extrapolated from standard curves of commercial shrimp TPM standard and corresponding in-house TPM standards are presented in Table 1.
Differences in TPM concentration of the same sample using different TPM standards differ from 40 to 600 times, which strongly indicates that the right choice of TPM standard is a critical step for accurate and precise determination of TPM concentration in seafood samples.
Извор:
XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts, 2023, 132-132Издавач:
- Beograd : Srpsko hemijsko društvo
Финансирање / пројекти:
- Imptox (An innovative analytical platform to investigate the effect and toxicity of micro and nano plastics combined with environmental contaminants on the risk of allergic disease in preclinical and clinical) (EU-H2020-965173)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200168 (Универзитет у Београду, Хемијски факултет) (RS-MESTD-inst-2020-200168)
- Serbian Academy of Sciences and Arts, grant number F-26
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - CONF AU - Krstić-Ristivojević, Maja AU - Jovanović, Vesna B. AU - Radomirović, Mirjana Ž. AU - Trifunović, Olga AU - Stanić-Vučinić, Dragana AU - Ćirković Veličković, Tanja PY - 2023 UR - http://cherry.chem.bg.ac.rs/handle/123456789/6024 AB - In the last 50 years, the annual per capita consumption of seafood products worldwide has more than doubled, from almost 10 kg in 1960 to over 20 kg in 2014. Seafood protein is an essential part of the diet in many countries, particularly where total protein intake is low [1]. However, as defined by the European Community, fish, and shellfish tropomyosins (TPM) are major allergens and major causes of anaphylaxis [2]. The increasing prevalence of food allergies is consistent with the increasing pollution of soil and water with plastic particles. To investigate the potential link between increasing plastic pollution and increasing food allergy prevalence, we aim to develop methods for precise and accurate monitoring of allergens and plastic in real seafood samples. TPM was isolated from shrimp (Litopenaeus vannamei), clams (Venerupis philippinarum), and mussels (Mytilus galloprovincialis). The obtained in-house TPM proteins from three different sources were resolved using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The concentration of TPM in seashell samples from different geographical origin was determined using a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with prior optimization of adequate TPM standard curve using commercial and non-commercial in-house prepared TPM standards. TPM standards resolved via 2D-PAGE revealed the presence of two isoforms of shrimp and mussels TPM standard, one dominant and one less abundant isoform. Two isoforms from both seafood sources, shrimp and mussels, are slightly different in molecular weight and pI value. As for the TPM standard obtained from clams, the 2D electrophoregram showed possibly eight isoforms with small differences in mass and pI values. Furthermore, the presence of three dominant isoforms can be observed that differ slightly in molecular mass, while other isoforms also differ in pI value. The ELISA results, regarding TPM standard curve optimization, showed that in both the commercial shrimp TPM and in-house shrimp TPM standards, sigmoidal concentration dependence is present in a range of 50 to 0.05 ng/ml, using serial double dilutions. On the other hand, TPM standards isolated from mussels and clams show sigmoidal concentration dependence in the range of 45 to 0.044 μg/ml with using the identical combination of capture and detection antibodies and serial double dilutions. TPM concentrations in clams and mussel samples extrapolated from standard curves of commercial shrimp TPM standard and corresponding in-house TPM standards are presented in Table 1. Differences in TPM concentration of the same sample using different TPM standards differ from 40 to 600 times, which strongly indicates that the right choice of TPM standard is a critical step for accurate and precise determination of TPM concentration in seafood samples. PB - Beograd : Srpsko hemijsko društvo C3 - XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts T1 - Tropomyosin quantification in seafood samples-right choice of standard makes a difference SP - 132 EP - 132 UR - https://hdl.handle.net/21.15107/rcub_cherry_6024 ER -
@conference{ author = "Krstić-Ristivojević, Maja and Jovanović, Vesna B. and Radomirović, Mirjana Ž. and Trifunović, Olga and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja", year = "2023", abstract = "In the last 50 years, the annual per capita consumption of seafood products worldwide has more than doubled, from almost 10 kg in 1960 to over 20 kg in 2014. Seafood protein is an essential part of the diet in many countries, particularly where total protein intake is low [1]. However, as defined by the European Community, fish, and shellfish tropomyosins (TPM) are major allergens and major causes of anaphylaxis [2]. The increasing prevalence of food allergies is consistent with the increasing pollution of soil and water with plastic particles. To investigate the potential link between increasing plastic pollution and increasing food allergy prevalence, we aim to develop methods for precise and accurate monitoring of allergens and plastic in real seafood samples. TPM was isolated from shrimp (Litopenaeus vannamei), clams (Venerupis philippinarum), and mussels (Mytilus galloprovincialis). The obtained in-house TPM proteins from three different sources were resolved using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The concentration of TPM in seashell samples from different geographical origin was determined using a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with prior optimization of adequate TPM standard curve using commercial and non-commercial in-house prepared TPM standards. TPM standards resolved via 2D-PAGE revealed the presence of two isoforms of shrimp and mussels TPM standard, one dominant and one less abundant isoform. Two isoforms from both seafood sources, shrimp and mussels, are slightly different in molecular weight and pI value. As for the TPM standard obtained from clams, the 2D electrophoregram showed possibly eight isoforms with small differences in mass and pI values. Furthermore, the presence of three dominant isoforms can be observed that differ slightly in molecular mass, while other isoforms also differ in pI value. The ELISA results, regarding TPM standard curve optimization, showed that in both the commercial shrimp TPM and in-house shrimp TPM standards, sigmoidal concentration dependence is present in a range of 50 to 0.05 ng/ml, using serial double dilutions. On the other hand, TPM standards isolated from mussels and clams show sigmoidal concentration dependence in the range of 45 to 0.044 μg/ml with using the identical combination of capture and detection antibodies and serial double dilutions. TPM concentrations in clams and mussel samples extrapolated from standard curves of commercial shrimp TPM standard and corresponding in-house TPM standards are presented in Table 1. Differences in TPM concentration of the same sample using different TPM standards differ from 40 to 600 times, which strongly indicates that the right choice of TPM standard is a critical step for accurate and precise determination of TPM concentration in seafood samples.", publisher = "Beograd : Srpsko hemijsko društvo", journal = "XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts", title = "Tropomyosin quantification in seafood samples-right choice of standard makes a difference", pages = "132-132", url = "https://hdl.handle.net/21.15107/rcub_cherry_6024" }
Krstić-Ristivojević, M., Jovanović, V. B., Radomirović, M. Ž., Trifunović, O., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2023). Tropomyosin quantification in seafood samples-right choice of standard makes a difference. in XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts Beograd : Srpsko hemijsko društvo., 132-132. https://hdl.handle.net/21.15107/rcub_cherry_6024
Krstić-Ristivojević M, Jovanović VB, Radomirović MŽ, Trifunović O, Stanić-Vučinić D, Ćirković Veličković T. Tropomyosin quantification in seafood samples-right choice of standard makes a difference. in XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts. 2023;:132-132. https://hdl.handle.net/21.15107/rcub_cherry_6024 .
Krstić-Ristivojević, Maja, Jovanović, Vesna B., Radomirović, Mirjana Ž., Trifunović, Olga, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Tropomyosin quantification in seafood samples-right choice of standard makes a difference" in XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts (2023):132-132, https://hdl.handle.net/21.15107/rcub_cherry_6024 .