Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human Serum
Folic acid (FA) is an important vitamin for human growth, especially for pregnant women. FA deficiency is associated with megaloblastic anemia, neural tube defects, cardiovascular diseases, irritability, diarrhea, and psychiatric disorders. Normally, FA molecules bind to folate-binding protein (FBP)...
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Journal of Analytical Methods in Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2019/5402903 |
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| author | Xiang Chen Qiyang Zhou Ting Zhang ChunXin Wang Zheng Yu Hadji Ahamada Zhonghu Bai Xuan Huang |
| author_facet | Xiang Chen Qiyang Zhou Ting Zhang ChunXin Wang Zheng Yu Hadji Ahamada Zhonghu Bai Xuan Huang |
| author_sort | Xiang Chen |
| collection | DOAJ |
| description | Folic acid (FA) is an important vitamin for human growth, especially for pregnant women. FA deficiency is associated with megaloblastic anemia, neural tube defects, cardiovascular diseases, irritability, diarrhea, and psychiatric disorders. Normally, FA molecules bind to folate-binding protein (FBP) in the serum as complex. Before quantify the FA concentration, a releasing procedure should be conducted. Alkaline condition and tris(2-carboxyethyl)phosphine (TCEP) are used to release binding FA to freeing state. In this work, a chemiluminescence immunoassay (CLIA) for human serum FA was established by competition model. Streptavidin (SA) was labeled to magnetic beads by an 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDAC/NHS) method. Activated biotin molecules were labeled to FBP molecules purified from milk. FA was labeled to horseradish peroxidase (HRP) by EDAC to activate the FA molecules. The pretreated samples or standards were added into the reaction tube with biotin-FBP and FA-horseradish peroxidase (HRP), FA in the sample compete with FA-HRP for binding to biotin-FBP, the signal is inversely proportional to the FA concentration. The method established shows good thermostability and performance. The limitation of detection (LOD) is 0.44 ng/mL. The intra-assay coefficient of variation (CV) is 3.6%–7.1%, the interassay CV is 4.2%–7.5%, and the recovery rate is 92.1%–103.5%. Cross reactivity (CR) was remarkably low with aminopterin, folinic acid, and methotrexate. The method shows good correlation with the FA CLIA product from Beckman Coulter; the equation is y = 0.9618x−0.1434 while the R2 value is 0.9224. The established method is sensitive, rapid, and accurate which can fully satisfy for the clinical requirement. |
| format | Article |
| id | doaj-art-8567ef9259b54198963187bb238b9280 |
| institution | Kabale University |
| issn | 2090-8865 2090-8873 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Analytical Methods in Chemistry |
| spelling | doaj-art-8567ef9259b54198963187bb238b92802025-02-03T05:53:31ZengWileyJournal of Analytical Methods in Chemistry2090-88652090-88732019-01-01201910.1155/2019/54029035402903Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human SerumXiang Chen0Qiyang Zhou1Ting Zhang2ChunXin Wang3Zheng Yu4Hadji Ahamada5Zhonghu Bai6Xuan Huang7School of Biotechnology, Jiangnan University, Wuxi, ChinaJiangsu Testing and Inspection Institute for Medical Devices, Nanjing, ChinaThe Affiliated Wuxi Maternity and Child Health Care Hospital of Nanjing Medical University, Wuxi, ChinaMedical Laboratory, Wuxi People Hospital Affiliated to Nanjing Medical University, Wuxi, ChinaSchool of Biotechnology, Jiangnan University, Wuxi, ChinaHematology and Clinical Biochemistry Department, Hospital EL-Maarouf, Moroni, ComorosSchool of Biotechnology, Jiangnan University, Wuxi, ChinaDepartment of Laboratory Medicine, Affiliated Hospital of Jiangnan University, Wuxi, ChinaFolic acid (FA) is an important vitamin for human growth, especially for pregnant women. FA deficiency is associated with megaloblastic anemia, neural tube defects, cardiovascular diseases, irritability, diarrhea, and psychiatric disorders. Normally, FA molecules bind to folate-binding protein (FBP) in the serum as complex. Before quantify the FA concentration, a releasing procedure should be conducted. Alkaline condition and tris(2-carboxyethyl)phosphine (TCEP) are used to release binding FA to freeing state. In this work, a chemiluminescence immunoassay (CLIA) for human serum FA was established by competition model. Streptavidin (SA) was labeled to magnetic beads by an 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDAC/NHS) method. Activated biotin molecules were labeled to FBP molecules purified from milk. FA was labeled to horseradish peroxidase (HRP) by EDAC to activate the FA molecules. The pretreated samples or standards were added into the reaction tube with biotin-FBP and FA-horseradish peroxidase (HRP), FA in the sample compete with FA-HRP for binding to biotin-FBP, the signal is inversely proportional to the FA concentration. The method established shows good thermostability and performance. The limitation of detection (LOD) is 0.44 ng/mL. The intra-assay coefficient of variation (CV) is 3.6%–7.1%, the interassay CV is 4.2%–7.5%, and the recovery rate is 92.1%–103.5%. Cross reactivity (CR) was remarkably low with aminopterin, folinic acid, and methotrexate. The method shows good correlation with the FA CLIA product from Beckman Coulter; the equation is y = 0.9618x−0.1434 while the R2 value is 0.9224. The established method is sensitive, rapid, and accurate which can fully satisfy for the clinical requirement.http://dx.doi.org/10.1155/2019/5402903 |
| spellingShingle | Xiang Chen Qiyang Zhou Ting Zhang ChunXin Wang Zheng Yu Hadji Ahamada Zhonghu Bai Xuan Huang Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human Serum Journal of Analytical Methods in Chemistry |
| title | Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human Serum |
| title_full | Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human Serum |
| title_fullStr | Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human Serum |
| title_full_unstemmed | Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human Serum |
| title_short | Development of a Sensitive Chemiluminescence Immunoassay for the Quantification of Folic Acid in Human Serum |
| title_sort | development of a sensitive chemiluminescence immunoassay for the quantification of folic acid in human serum |
| url | http://dx.doi.org/10.1155/2019/5402903 |
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