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| Development and Characterization of Monoclonal Antibodies Against Human Heart Type Fatty Acid Binding Protein and Its Application in Lateral Flow Immunoassay |
| KANG Ke-ren1,2, WU Pei-dian2, HUANG Qi-lin2, LI Yue-lin2, LI Gao-hui2, TANG Shi-xing2, WANG Ji-hua2 |
1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China;
2. National Engineering Laboratory of Rapid Diagnostic Test. Guangzhou Wondfo Biotech Co. Ltd., Guangzhou 510663, China |
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Abstract AIM: To develop and characterize monoclonal antibodies (mAb) against human heart type fatty acid binding protein (H-FABP); To develop a rapid lateral flow immunoassay for detecting H-FABP in plasma using the identified mAbs. Methods: BALB/c mice were immunized with H-FABP. The hybridoma cells were generated by fusing spleen cells and myeloma cells SP2/0 followed by screening and clonally selecting. The mAbs were purified from ascites and characterized by indirect ELISA and Western Blot, and were used for developing a rapid lateral flow immunoassay.Results: Twelve hybridoma cell lines were found to secret anti-H-FABP with high titers, specificity and affinity. Among them, the antibody pair of 3D1 and 5F4 were used as conjugate and coating antibody, respectively, in lateral flow immunoassay. The newly developed assay was used to detect 203 patients plasma. A 100% consistence was observed when compared with the reference kit.Conclusion: Two mAbs against H-FABP with high titers, specificity and affinity were obtained. The rapid lateral flow immunoassay was successfully developed for sensitive and specific detection of H-FABP in human plasma by using these mAbs.
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Received: 28 October 2012
Published: 25 January 2013
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