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Advanced Pharmaceutical Bulletin
ISSN: 2228-5881      eISSN: 2251-7308  
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Article History
Submitted: 16 Jul 2014
Revised: 09 Sep 2014
First published online: 05 Mar 2015

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Adv Pharm Bull. 2015;5(1):97-102 doi: 10.5681/apb.2015.013
PMID:25789225        PMCID:PMC4352229

Large Scale Generation and Characterization of Anti-Human IgA Monoclonal Antibody in Ascitic Fluid of Balb/c Mice

Original Research

Fatemeh Ezzatifar 1,2,3, Jafar Majidi 1,2,4 * , Behzad Baradaran 2,4, Leili Aghebati Maleki 2,4, Jalal Abdolalizadeh 1,2, Mehdi Yousefi 2,4

1 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
2 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
3 Tabriz University of Medical Sciences, International Branch Aras, Tabriz University of Medical Sciences, Tabriz, Iran.
4 Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.



Abstract
Purpose: Monoclonal antibodies are potentially powerful tools used in biomedical research, diagnosis, and treatment of infectious diseases and cancers. The monoclonal antibody against Human IgA can be used as a diagnostic application to detect infectious diseases. The aim of this study was to improve an appropriate protocol for large-scale production of mAbs against IgA. Methods: For large-scale production of the monoclonal antibody, hybridoma cells that produce monoclonal antibodies against Human IgA were injected intraperitoneally into Balb/c mice that were previously primed with 0.5 ml Pristane. After ten days, ascitic fluid was harvested from the peritoneum of each mouse. The ELISA method was carried out for evaluation of the titration of produced mAbs. The ascitic fluid was investigated in terms of class and subclass by a mouse mAb isotyping kit. MAb was purified from the ascitic fluid by ion exchange chromatography. The purity of the monoclonal antibody was confirmed by SDS-PAGE, and the purified monoclonal antibody was conjugated with HRP. Results: Monoclonal antibodies with high specificity and sensitivity against Human IgA were prepared by hybridoma technology. The subclass of antibody was IgG1 and its light chain was the kappa type. Conclusion: This conjugated monoclonal antibody could have applications in designing ELISA kits in order to diagnose different infectious diseases such as toxoplasmosis and H. Pylori.





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