Biotechnology & nanotechnology
Ezzat Nourizadeh
Volume 28, Issue 5 , November and December 2021, , Pages 775-789
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 ...
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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.
immunology & Biochemistry
Ezzat Nourizadeh
Volume 28, Issue 2 , May and June 2021, , Pages 302-310
Abstract
Introduction: Leishmania parasitic infections are the important causes of health problems in many parts of the world, especially in developing countries. Monoclonal antibodies have been used as valuable tools for the detection, treatment and characterization of the antigenic markers of parasites. This ...
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Introduction: Leishmania parasitic infections are the important causes of health problems in many parts of the world, especially in developing countries. Monoclonal antibodies have been used as valuable tools for the detection, treatment and characterization of the antigenic markers of parasites. This study through applicable techniques aims to produce mAbs against Iranian type of Leishmania( L). infantum. Materials and Methods: Standard strains were cultured and their antigens were used. BALB/c mice were injected with freeze-thawed promastigote twice together with Freund adjuvant. Three days before cell-fusion, the antigen through vein was injected into the mice. Then the mice were killed. After that their spleen lymphocytes were mingled with myeloma SP2/0. In the next step, the isolation of monoclones was performed by limiting dilution method. Results: 16 mAb against promastigote form of L.infantum parasite were obtained ,3 of which showed optical density (OD) more than 1nm, designated as 5D2 FV, 4G5 FV and 5D6 FIV. Then, anti-promastigotes L.infantum mAbs were obtained from these hybrids. These antibodies are effective in the logarithmic phase of the parasite. Conclusion: It seams these antibodies can demonstrate reaction against Iranian strain of promastigotes L. infantum and can be employed in the diagnosis of kalazar disease