Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic Immunoassay
Serum ferritin (SF) is an iron-rich protein tightly connected with iron homeostasis, and the variations are frequently observed in diseased states, including iron-deficiency anemia, inflammation, liver disease, and tumors, which renders SF level an indicator of potential malignancies in clinical pra...
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| Language: | English |
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MDPI AG
2022-08-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/12/8/1080 |
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| author | Yaozhong Hu Jing Lin Yi Wang Sihao Wu Jing Wu Huan Lv Xuemeng Ji Serge Muyldermans Yan Zhang Shuo Wang |
| author_facet | Yaozhong Hu Jing Lin Yi Wang Sihao Wu Jing Wu Huan Lv Xuemeng Ji Serge Muyldermans Yan Zhang Shuo Wang |
| author_sort | Yaozhong Hu |
| collection | DOAJ |
| description | Serum ferritin (SF) is an iron-rich protein tightly connected with iron homeostasis, and the variations are frequently observed in diseased states, including iron-deficiency anemia, inflammation, liver disease, and tumors, which renders SF level an indicator of potential malignancies in clinical practice. Nanobodies (Nbs) have been widely explored and developed into theranostic reagents. Surprisingly, no reports stated the identification of anti-SF Nbs, nor the potential of such Nbs as a diagnostic tool. In this study, we generated SF-specific Nbs and provided novel clinical diagnostic approaches to develop an immunoassay. An immune library was constructed after immunizing an alpaca with SF, and five Nbs specifically targeting human SF were retrieved. The obtained Nbs exhibited robust properties including high stability, affinity, and specificity. Then, an ELISA-based test using a heterologous Nb-pair was developed. The calibration curve demonstrated a linear range of SF between 9.0 to 1100 ng/mL, and a limit of detection (LOD) of 1.01 ng/mL. The detecting recovery and coefficient variation (CV) were determined by spiking different concentrations of SF into the serum sample, to verify the successful application of our selected Nbs for SF monitoring. In general, this study generated SF-specific Nbs and demonstrated their potential as diagnostic immunoassay tools. |
| first_indexed | 2024-03-09T11:49:46Z |
| format | Article |
| id | doaj.art-c11d14e709704a97a0b8c6339824e783 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T11:49:46Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-c11d14e709704a97a0b8c6339824e7832023-11-30T23:16:11ZengMDPI AGBiomolecules2218-273X2022-08-01128108010.3390/biom12081080Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic ImmunoassayYaozhong Hu0Jing Lin1Yi Wang2Sihao Wu3Jing Wu4Huan Lv5Xuemeng Ji6Serge Muyldermans7Yan Zhang8Shuo Wang9Research Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, 1050 Brussels, BelgiumResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaResearch Institute of Public Health, School of Medicine, Nankai University, Tianjin 300071, ChinaSerum ferritin (SF) is an iron-rich protein tightly connected with iron homeostasis, and the variations are frequently observed in diseased states, including iron-deficiency anemia, inflammation, liver disease, and tumors, which renders SF level an indicator of potential malignancies in clinical practice. Nanobodies (Nbs) have been widely explored and developed into theranostic reagents. Surprisingly, no reports stated the identification of anti-SF Nbs, nor the potential of such Nbs as a diagnostic tool. In this study, we generated SF-specific Nbs and provided novel clinical diagnostic approaches to develop an immunoassay. An immune library was constructed after immunizing an alpaca with SF, and five Nbs specifically targeting human SF were retrieved. The obtained Nbs exhibited robust properties including high stability, affinity, and specificity. Then, an ELISA-based test using a heterologous Nb-pair was developed. The calibration curve demonstrated a linear range of SF between 9.0 to 1100 ng/mL, and a limit of detection (LOD) of 1.01 ng/mL. The detecting recovery and coefficient variation (CV) were determined by spiking different concentrations of SF into the serum sample, to verify the successful application of our selected Nbs for SF monitoring. In general, this study generated SF-specific Nbs and demonstrated their potential as diagnostic immunoassay tools.https://www.mdpi.com/2218-273X/12/8/1080serum ferritinnanobodydiagnosisimmunoassayclinical indicator |
| spellingShingle | Yaozhong Hu Jing Lin Yi Wang Sihao Wu Jing Wu Huan Lv Xuemeng Ji Serge Muyldermans Yan Zhang Shuo Wang Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic Immunoassay Biomolecules serum ferritin nanobody diagnosis immunoassay clinical indicator |
| title | Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic Immunoassay |
| title_full | Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic Immunoassay |
| title_fullStr | Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic Immunoassay |
| title_full_unstemmed | Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic Immunoassay |
| title_short | Identification of Serum Ferritin-Specific Nanobodies and Development towards a Diagnostic Immunoassay |
| title_sort | identification of serum ferritin specific nanobodies and development towards a diagnostic immunoassay |
| topic | serum ferritin nanobody diagnosis immunoassay clinical indicator |
| url | https://www.mdpi.com/2218-273X/12/8/1080 |
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