Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy products
Abstract Aflatoxin M1 (AFM1) is known to be carcinogenic, mutagenic, and teratogenic and poses a serious threat to food safety and human health, which makes its surveillance critical. In this study, an indirect competitive ELISA (icELISA) based on a nanobody (Nb M4) was developed for the sensitive a...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-024-83869-4 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841559662940389376 |
|---|---|
| author | Li Yi Haiyuan Liu Yingda Liu Aiyisi Jing He Liang Ming Jirimutu |
| author_facet | Li Yi Haiyuan Liu Yingda Liu Aiyisi Jing He Liang Ming Jirimutu |
| author_sort | Li Yi |
| collection | DOAJ |
| description | Abstract Aflatoxin M1 (AFM1) is known to be carcinogenic, mutagenic, and teratogenic and poses a serious threat to food safety and human health, which makes its surveillance critical. In this study, an indirect competitive ELISA (icELISA) based on a nanobody (Nb M4) was developed for the sensitive and rapid detection of AFM1 in dairy products. In our previous work, Nb M4 was screened from a Bactrian-camel-immunized phage-displayed library. It exhibits VH-like features, possesses higher thermal stability than monoclonal antibody (mAb 1E6) and tightly binds to AFM1–BSA with a K D value of 2.5 nM. Under the optimal conditions, its half-maximal inhibitory concentration was 0.338 ng/mL, the limit of detection was 0.051 ng/mL, and linearity was noted in the range of 0.168–0.679 ng/mL. Nb M4 displayed almost no cross-reactivity with other mycotoxins. No matrix effect was observed in milk and milk powder samples, and the matrix effect in yogurt samples could be weakened by 2-fold dilution. Furthermore, validation studies in spiked samples (milk, yogurt, and milk powder) resulted in good recoveries of 95.40–111.33%, with a low coefficient of variation (2.89–6.78%). High-performance liquid chromatography was used to evaluate the accuracy and reliability of the developed icELISA, which indicated a satisfactory consistent correlation (R2 = 0.9722). This study highlights the potential of Nb M4 as a promising component for detecting AFM1 in dairy products. |
| format | Article |
| id | doaj-art-665ec5455f7b4a9981216d904f177912 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-665ec5455f7b4a9981216d904f1779122025-01-05T12:17:47ZengNature PortfolioScientific Reports2045-23222025-01-0115111510.1038/s41598-024-83869-4Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy productsLi Yi0Haiyuan Liu1Yingda Liu2Aiyisi3Jing He4Liang Ming5Jirimutu6Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, College of Food Science and Engineering, Inner Mongolia Agricultural UniversityKey Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, College of Food Science and Engineering, Inner Mongolia Agricultural UniversityKey Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, College of Food Science and Engineering, Inner Mongolia Agricultural UniversityInner Mongolia Autonomous Region International Mongolian HospitalKey Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, College of Food Science and Engineering, Inner Mongolia Agricultural UniversityKey Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, College of Food Science and Engineering, Inner Mongolia Agricultural UniversityKey Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, College of Food Science and Engineering, Inner Mongolia Agricultural UniversityAbstract Aflatoxin M1 (AFM1) is known to be carcinogenic, mutagenic, and teratogenic and poses a serious threat to food safety and human health, which makes its surveillance critical. In this study, an indirect competitive ELISA (icELISA) based on a nanobody (Nb M4) was developed for the sensitive and rapid detection of AFM1 in dairy products. In our previous work, Nb M4 was screened from a Bactrian-camel-immunized phage-displayed library. It exhibits VH-like features, possesses higher thermal stability than monoclonal antibody (mAb 1E6) and tightly binds to AFM1–BSA with a K D value of 2.5 nM. Under the optimal conditions, its half-maximal inhibitory concentration was 0.338 ng/mL, the limit of detection was 0.051 ng/mL, and linearity was noted in the range of 0.168–0.679 ng/mL. Nb M4 displayed almost no cross-reactivity with other mycotoxins. No matrix effect was observed in milk and milk powder samples, and the matrix effect in yogurt samples could be weakened by 2-fold dilution. Furthermore, validation studies in spiked samples (milk, yogurt, and milk powder) resulted in good recoveries of 95.40–111.33%, with a low coefficient of variation (2.89–6.78%). High-performance liquid chromatography was used to evaluate the accuracy and reliability of the developed icELISA, which indicated a satisfactory consistent correlation (R2 = 0.9722). This study highlights the potential of Nb M4 as a promising component for detecting AFM1 in dairy products.https://doi.org/10.1038/s41598-024-83869-4Aflatoxin M1Indirect competitive ELISANanobodyDairy products |
| spellingShingle | Li Yi Haiyuan Liu Yingda Liu Aiyisi Jing He Liang Ming Jirimutu Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy products Scientific Reports Aflatoxin M1 Indirect competitive ELISA Nanobody Dairy products |
| title | Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy products |
| title_full | Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy products |
| title_fullStr | Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy products |
| title_full_unstemmed | Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy products |
| title_short | Nanobody-based indirect competitive ELISA for the detection of aflatoxin M1 in dairy products |
| title_sort | nanobody based indirect competitive elisa for the detection of aflatoxin m1 in dairy products |
| topic | Aflatoxin M1 Indirect competitive ELISA Nanobody Dairy products |
| url | https://doi.org/10.1038/s41598-024-83869-4 |
| work_keys_str_mv | AT liyi nanobodybasedindirectcompetitiveelisaforthedetectionofaflatoxinm1indairyproducts AT haiyuanliu nanobodybasedindirectcompetitiveelisaforthedetectionofaflatoxinm1indairyproducts AT yingdaliu nanobodybasedindirectcompetitiveelisaforthedetectionofaflatoxinm1indairyproducts AT aiyisi nanobodybasedindirectcompetitiveelisaforthedetectionofaflatoxinm1indairyproducts AT jinghe nanobodybasedindirectcompetitiveelisaforthedetectionofaflatoxinm1indairyproducts AT liangming nanobodybasedindirectcompetitiveelisaforthedetectionofaflatoxinm1indairyproducts AT jirimutu nanobodybasedindirectcompetitiveelisaforthedetectionofaflatoxinm1indairyproducts |