Document Type : Original Article
Author
Assistant Professor, Department of Microbiology, College of Science, University of Mohaghegh Ardabili, Ardabil, Iran
Abstract
Introduction: Leishmania (L.) infantum is the etiologic cause of visceral leishmaniasis in Iran. Efficient vaccines and diagnosis methods are required to control leishmaniasis. The aim of this study is produce and optimize monoclonal antibodies against promastigotes forms of L. infantum antigen.
Materials and Methods: The mice were vaccinated with the L. infantum antigen and their antibody titers were determined by the ELISA method. Spleen cells of the most immune mouse were fused with SP2/0 in the presence of Poly Ethylene Glycol.The effect of supernatant of SP2/0 and mice peritoneum macrophage cells culture (SSMCC) on hybridoma cell proliferation was studied.
Results: Among the 12 fusion, a total of 26 monoclonal were positive.12 of which had acceptable optical absorbance in OD 450 nm. Finally, 4 clones, designated as 8D2 FVI6, 8D2 FVI3, 6G2 FV4 and 6G2 FV3. From these hybrids, anti-promastigotes L. infantum monoclonal antibodies were obtained. SSMCC was shown to play a key role in hybridoma proliferation and of mAb production. It seemed that SSMCC is rich of growth factors.
Conclusion: It seems in the near future, this SCCSM can be used as a growth factor for cancerous and non-cancerous cells in research centers at a wider level.
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