Affiliations: [a] Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
| [b] Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
| [c] Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
Correspondence:
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Corresponding author: Jafar Majidi, Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. Tel.: +98 411 3364665; Fax: +98 411 3364665; E-mail: [email protected].
Abstract: Objective:Immunoglobulin A (IgA) is the second immunoglobulin in human serum. Monoclonal antibodies have many potential uses in diagnosis, treatment and purification. The conjugated monoclonal antibodies against human IgA are utilized in diagnostic kits. Methods:For production of monoclonal antibodies against human Immunoglobulin A, spleen cells of the immune mouse were fused with SP2/0 as myeloma cells and Poly Ethylene Glycol (PEG). Supernatant of hybridoma cells was screened to determine the antibody by ELISA. The appropriate clones were selected for limiting dilution (L.D). Ultimately, appropriate monoclone was injected intraperitoneally into the mouse that has been primed with Pristane. Results:In current study, 175 clones were obtained of which 5 clones had absorbance values of about 3 were selected for limiting dilution. Between these clones, 3-D5monoclone with IgG1 subclass was selected as appropriate one and it was reproduced in FCS free RPMI 1640. For large scale production in invivo, the appropriate clone was implanted in the peritoneum of the Balb/c mouse and its titer was determined, which showed 1/100,000 dilution for ascitic fluid, having no cross reactivity with IgM & IgG. Conclusions:Monoclonal antibody was purified by chromatography, confirmed by SDS-PAGE and then conjugated with enzyme and used for diagnostic kits.
Keywords: Production, monoclonal antibody, ELISA, limiting dilution, IgA
