Faculty of Paramedicine - Jalal Abdolalizadeh

Jalal Abdolalizadeh

Laboratory Sciences

Academic Position : Associate Professor

Start service : 2012

Section : Laboratory Sciences

Educational Background :

Phone : 33371971

Email Address : jabdolalizadeh@gmail.com

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Article Title : A unique report: Development of super Anti-Human IgG Monoclone with Optical density over than 3

Master Auther : Jafae Majidi

Journal Title and Number : Advanced pharmaceutical Bulletine 3

Authers : Leili Aghebati Maleki - Behzad Baradaran - Jalal Abdolalizadeh - Fatemeh Ezzatifar - Jafar Majidi

Publish : 2013

summary :

Article Title : Development and characterization of monoclonal antibodies against human CD20 in Balb/c mice

Master Auther :

Journal Title and Number : Human Antibodies 21 (2012) 57–64

Authers : Koushan Sineh Sepehra,b, Behzad Baradarana,b,∗, Jafar Majidib,c, Jalal Abdolalizadehb,c, Leili Aghebatib and Fatemeh Zare Shahnehb

Publish : 2012

summary : Abstract. BACKGROUND: Targeting CD20 antigen on B-lymphocytes provides good opportunity for management of the target cells in patients with B-cell malignancies. By the advent of hybridoma technology, monoclonal antibodies applications exert extensive changes in medical fields such as diagnosis, treatment and purification. OBJECTIVE: The prim aim of this study was to produce monoclonal antibody against CD20 for exploitation in diagnosis. METHODS: In this study, Balb/c mice were immunized with two peptides from extracellular domain of CD20. Poly Ethylene Glycol (PEG) fused spleen cells of the most immune mouse with SP2/0 (myeloma cells). Supernatant of hybridoma cells were screened for detection of antibody by ELISA. The desired clones were selected for limiting dilution (L.D). Afterward, specificity and cross reactivity of these antibodies were determined by immunological assay such as ELISA and western blot analysis (WB) and Immunofluorescence. Large scale of monoclonal antibodies was produced by ascetic fluid method. Monoclonal antibody was purified by chromatography then confirmed by SDS-PAGE. RESULTS: In this study, between five positive clone wells, 3 clones were chosen for limiting dilution. Limiting dilution product was one monoclone with absorbance about 2. CONCLUSIONS: These results indicate that such monoclonal antibodies against CD20 can be used in diagnosis of CD20 in the cells surface.

Article Title : Cloning and Expression of Leishmania infantum P4 gene for production of recombinant P4 antigen

Master Auther : Farajnia S.

Journal Title and Number : Pharmaceutical Sciences, Spring (2008) 53-58

Authers : Farajnia S.1,2, Beh-Pajooh A. 2, Alimohammadian M.H. 4, Babaei H1, 3., Asgharzadeh M. 5, Majidi J. 6, Kazemi A. 6, Mesgari M. 1, Mohammadnejad L. 2, Abdolalizadeh J.

Publish : 2008

summary : Visceral leishmaniasis (VL) or Kala-azar caused by Leishmania infantum is a severe endemic disease in the Mediterranean basin countries including Iran. The Drugs available for treatment of VL are toxic and drug resistance is increasing in many parts of the world, thus, it is believed that vaccine development is an ideal method for prevention and control of VL. The aim of this study was to prepare recombinant P4 protein from Leishmania infantum and evaluate its immunogenicity in VL patients. Methods: DNA was extracted from Leishmania infantum and used for amplification of P4 gene by PCR. The PCR product was cloned, sequenced and expressed in E. coli using pET 28a expression vector. Recombinant P4 protein were purified and used for analysis of sera of VL patients. Results: Analysis of the sequence of Leishmania infantum P4 gene (Li-P4) revealed that the gene consists of an ORF of 951bp with a 89% homology with P4 gene of cutaneous leishmaniasis agents. Expression of Li-P4 gene in E. coli resulted in high levels of recombinant proteins with molecular weight of 33 KD in SDS-PAGE. Immunoblotting analysis of the purified Li-P4 with sera of VL patients indicated that Li-P4 protein is a highly immunogenic protein expressed in the amastigote-stage of Leishmania infantum. Conclusion: This is the first study on isolation and characterization of P4 antigen from Leishmania infantum and indicates that Li-P4 protein is highly immunogenic and could be considered as a potent vaccine candidate against VL caused by Leishmania infantum. Keywords: Visceral leishmaniasis, P4 antigen, Leishmania infantum.

Article Title : Production and purification of polyclonal antibody against human kappa light chain. Journal of Biological Sciences.

Master Auther :

Journal Title and Number : Journal of Biological Sciences. 2008; 8(3): 683-686.

Authers : Abdolalizadeh J, Majidi J, Farajnia S

Publish : 2008

summary :

Article Title : Long-term exercise training affects age-induced changes in HSP70

Master Auther :

Journal Title and Number : Gen. Physiol. Biophys. (2008), 27, 263–270

Authers : Farhad G. Soufi1,2, Safar Farajnia1,3, Naser Aslanabadi1,4, Naser Ahmadiasl, Mohammadreza Alipour, Mohsen Alipour, Yousef Doustar, Jalal Abdolalizadeh and Farzam Sheikhzadeh

Publish : 2008

summary : The aim of this study was to test the effects of age and long-term exercise training on antioxidant, heat shock protein 70 (HSP70) expression and apoptosis by comparing the hearts of sedentary and trained rats. Training groups went under 3-, 6- and 9-months of regular exercise (25 m/min with a 0% slope, 60 min/day and 6 days/week). Level of glutathione increased with age in trained and sedentary control rats but level of this factor unchanged by training. Activity of mitochondrial superoxide dismutase (mtSOD) increased in heart homogenates of 6- and 9-months trained animals as compared with their sedentary. The rates of apoptosis were increased with age but level of apoptosis in 9-months trained group was significantly lower than corresponding sedentary. Levels of HSP70 expression were significantly decreased with age while long-term training induced marked increase in HSP70 expression levels. These results show that a long-term regular exercise affects age-induced changes in mtSOD, HSP70 and apoptosis as it increases mtSOD activities and HSP70 expression levels and elicits a marked reduction in apoptosis rate in rat myocardium. However, a shorter training program is not effective.

Article Title : Development and characterization of monoclonal antibodies against human CD20 in Balb/c mice

Master Auther :

Journal Title and Number : Hum Antibodies. 2012;21(3-4):57-64. doi: 10.3233/HAB-130263

Authers : Sepehr KS, Baradaran B, Majidi J, Abdolalizadeh J, Aghebati L, Shahneh FZ.

Publish : 2012

summary : Targeting CD20 antigen on B-lymphocytes provides good opportunity for management of the target cells in patients with B-cell malignancies. By the advent of hybridoma technology, monoclonal antibodies applications exert extensive changes in medical fields such as diagnosis, treatment and purification. OBJECTIVE: The prim aim of this study was to produce monoclonal antibody against CD20 for exploitation in diagnosis. METHODS: In this study, Balb/c mice were immunized with two peptides from extracellular domain of CD20. Poly Ethylene Glycol (PEG) fused spleen cells of the most immune mouse with SP2/0 (myeloma cells). Supernatant of hybridoma cells were screened for detection of antibody by ELISA. The desired clones were selected for limiting dilution (L.D). Afterward, specificity and cross reactivity of these antibodies were determined by immunological assay such as ELISA and western blot analysis (WB) and Immunofluorescence. Large scale of monoclonal antibodies was produced by ascetic fluid method. Monoclonal antibody was purified by chromatography then confirmed by SDS-PAGE. RESULTS: In this study, between five positive clone wells, 3 clones were chosen for limiting dilution. Limiting dilution product was one monoclone with absorbance about 2. CONCLUSIONS: These results indicate that such monoclonal antibodies against CD20 can be used in diagnosis of CD20 in the cells surface.

Article Title : Target therapy of cancer: Implementation of monoclonal antibodies and nanobodies, Human Antibodies

Master Auther : Majidi J

Journal Title and Number : Human Antibodies 18 (2009) 81–100

Authers : Majidi J, Barar J, Baradaran B, Abdolalizadeh J and Omidi Y

Publish : 2009

summary : In the past decades, the mainstay of systemic therapy for solid and haematological malignancies was chemotherapy; nevertheless this modality has the drawbacks such as drug resistance and eliciting sever cytotoxicity in the normal tissue. To resolve such downsides, the cancer therapy modalities need to be advanced with more effective and tolerable treatments to specifically target the malignant cell with minimal adverse consequences. In fact, characteristically, the malignant diseases are self sufficiency in growth signals along with insensitivity to growth inhibition. They can also evade from apoptosis, have limitless replicative potential, induce angiogenesis and possess metastasis potential. Given that the most of these characteristics are often due to genetic defects, thus key to the development of targeted therapies is the ability to use such processes to phenotypically distinguish the tumor from its normal counterpart by its specific/selective markers. The therapeutic monoclonal antibodies (mAbs) are deemed to be a class of novel agents that can specifically target and disrupt molecular pathways underlying tumorigenesis. The mAbs are produced by a single clone of B-cells, and are monospecific and homogeneous. Since Kohler and Milstein heralded a new era in antibody research and clinical development by the discovery of hybridoma technology in 1975, more than 20 mAbs have been approved by the US Food and Drug Administration (FDA) for treatment of obdurate diseases, including different types of cancers. Mouse hybridomas were the first reliable source of monoclonal antibodies which were developed for several in vivo therapeutic applications. Accordingly, the recombinant antibodies have been reduced in size, rebuilt into multivalent molecules and fused with different moieties such as radionuclides, toxins and enzymes. The emergence of recombinant technologies, transgenic animals and phage display technology has revolutionized the selection, humanization and production of antibodies. This review focuses on implementation of the mAbs and nanobodies fragments for cancer therapy

Article Title : Designing of enzyme linked immunosorbent assay (ELISA) kit for diagnosis copro-antigens of Giardia lamblia, African

Master Auther : Barazesh A

Journal Title and Number : African Journal of Biotechnology Vol. 9(31), pp. 5025-5027, 2 August, 2010

Authers : Barazesh A, Majidi J, Fallah E, Jamali R, Abdolalizadeh J and Gholikhani R

Publish : 2010

summary : The sensitivity of microscopic examination of fecal samples to recognize Giardia parasites is low. In the methods based on antigen scanning of parasites such as enzyme linked immunosorbent assay (ELISA), copro-antigens of parasite will be traced and diagnosed even if the live parasite is absent in the fecal samples. To design this method, a pure antibody against parasite as well as an antibody conjugated to a proper enzyme is needed. In this study, an anti-Giardia IgG extracted from serum of contaminated rabbit was purified by ion-exchange chromatography and conjugated to the enzyme horse radish peroxidase (HRP). This antibody was used to design direct and indirect ELISA kits to measure conjugation titer. In both direct and indirect ELISA methods, optical densities (ODs) were 1 by using dilution of 1/4000 of conjugation. According to the results of both tests and the success in produced conjugate, it could be proceeded to prepare ELISA kits to diagnose giardiasis infections in various samples.

Article Title : Production and characterization of murine monoclonal antibody against synthetic peptide of CD34

Master Auther : Maleki LA

Journal Title and Number : Production and characterization of murine monoclonal antibody against synthetic peptide of CD34 .Human Antibodies. 1;22(1): 1-8

Authers : Maleki LA, Majidi J, Baradaran B, Abdolalizadeh J, Akbari AM

Publish : 2013

summary :

Article Title : Affinity purification of tumor necrosis factor-α expressed in Raji cells by produced scFv antibody coupled CNBr-activated sepharose

Master Auther : Omidi Y

Journal Title and Number : Advanced Pharmaceutical Bulletin. 2013; 3(1): 19-23.

Authers : Abdolalizadeh J, Majidi J, Nouri M, Baradaran B, Movassaghpour AA, Farajnia S and Omidi Y

Publish : 2013

summary : Purpose: Recombinant tumor necrosis factor-alpha (TNF-α) has been utilized as an antineoplastic agent for the treatment of patients with melanoma and sarcoma. It targets tumor cell antigens by impressing tumor-associated vessels. Protein purification with affinity chromatography has been widely used in the downstream processing of pharmaceutical-grade proteins. Methods: In this study, we examined the potential of our produced anti-TNF-scFv fragments for purification of TNF-α produced by Raji cells. he Raji cells were induced by lipopolysaccharides (LPS) to express TNF-α. Western blotting and Fluorescence-activated cell sorting (FACS) flow cytometry analyses were used to evaluate the TNF-α expression. The anti-TNF-α scFv selected from antibody phage display library was coupled to CNBr-activated sepharose 4B beads used for affinity purification of expressed TNF-α and the purity of the protein was assessed by SDS-PAGE. Results: Western blot and FACS flow cytometry analyses showed the successful expression of TNF-α with Raji cells. SDS-PAGE analysis showed the performance of scFv for purification of TNF-α protein with purity over 95%. Conclusion: These findings confirm not only the potential of the produced scFv antibody fragments but also this highly pure recombinant TNF-α protein can be applied for various in vitro and in vivo applications.

Article Title : Affinity purification of tumor necrosis factor-α expressed in Raji cells by produced scFv antibody coupled CNBr-activated sepharose

Master Auther : Omidi Y

Journal Title and Number : Advanced Pharmaceutical Bulletin. 2013; 3(1): 19-23.

Authers : Abdolalizadeh J, Majidi J, Nouri M, Baradaran B, Movassaghpour AA, Farajnia S and Omidi Y

Publish : 2013

summary : Purpose: Recombinant tumor necrosis factor-alpha (TNF-α) has been utilized as an antineoplastic agent for the treatment of patients with melanoma and sarcoma. It targets tumor cell antigens by impressing tumor-associated vessels. Protein purification with affinity chromatography has been widely used in the downstream processing of pharmaceutical-grade proteins. Methods: In this study, we examined the potential of our produced anti-TNF-scFv fragments for purification of TNF-α produced by Raji cells. he Raji cells were induced by lipopolysaccharides (LPS) to express TNF-α. Western blotting and Fluorescence-activated cell sorting (FACS) flow cytometry analyses were used to evaluate the TNF-α expression. The anti-TNF-α scFv selected from antibody phage display library was coupled to CNBr-activated sepharose 4B beads used for affinity purification of expressed TNF-α and the purity of the protein was assessed by SDS-PAGE. Results: Western blot and FACS flow cytometry analyses showed the successful expression of TNF-α with Raji cells. SDS-PAGE analysis showed the performance of scFv for purification of TNF-α protein with purity over 95%. Conclusion: These findings confirm not only the potential of the produced scFv antibody fragments but also this highly pure recombinant TNF-α protein can be applied for various in vitro and in vivo applications.

Article Title : Targeting cytokines: production and characterization of anti-TNF-α scFvs by phage display technology

Master Auther : Omidi Y

Journal Title and Number : Currentm Pharmaceutical Design. 2013; 19(15): 2839-47.

Authers : Abdolalizadeh J, Nouri M, Majidi J, Baradaran B, Barzegari A, Barar J and Omidi Y.

Publish : 2013

summary : The antibody display technology (ADT) such as phage display (PD) has substantially improved the production of monoclonal antibodies (mAbs) and Ab fragments through bypassing several limitations associated with the traditional approach of hybridoma technology. In the current study, we capitalized on the PD technology to produce high affinity single chain variable fragment (scFv) against tumor necrosis factor-alpha (TNF- ), which is a potent pro-inflammatory cytokine and plays important role in various inflammatory diseases and malignancies. To pursue production of scFv antibody fragments against human TNF- , we performed five rounds of biopanning using stepwise decreased amount of TNF- (1 to 0.1 g), a semi-synthetic phage antibody library (Tomlinson I + J) and TG1 cells. Antibody clones were isolated and selected through enzyme-linked immunosorbent assay (ELISA) screening. The selected scFv antibody fragments were further characterized by means of ELISA, PCR, restriction fragment length polymorphism (RFLP) and Western blot analyses as well as fluorescence microscopy and flow cytometry. Based upon binding affinity to TNF- , 15 clones were selected out of 50 positive clones enriched from PD in vitro selection. The selected scFvs displayed high specificity and binding affinity with Kd values at nm range to human TNF- . The immunofluorescence analysis revealed significant binding of the selected scFv antibody fragments to the Raji B lymphoblasts. The effectiveness of the selected scFv fragments was further validated by flow cytometry analysis in the lipopolysaccharide (LPS) treated mouse fibroblast L929 cells. Based upon these findings, we propose the selected fully human anti-TNF- scFv antibody fragments as potential immunotherapy agents that may be translated into preclinical/clinical applications.

Article Title : Inhibitory and Cytotoxic Activities of Salvia Officinalis L. Extract on Human Lymphoma and Leukemia Cells by Induction of Apoptosis

Master Auther : Behzad Baradaran

Journal Title and Number : Advanced Pharmaceutical Bulletin, 2013, 3(1), 51-55

Authers : Zare Shahneh F, Valiyari S, Baradaran B, Abdolalizadeh J, Bandehagh A, Azad Mehr A, Haji Aghaee

Publish : 2013

summary : Purpose: Salvia officinalis L., also known as Maryam Goli, is one of the native plants used to Persian medicinal herbs. Hence, the objective of this study was to examine the in vitro cytotoxic activities of a standardized crude methanol extracts prepared from Salvia officinalis L., on a non-Hodgkin’s B-cell lymphoma (Raji) and human leukemic monocyte lymphoma (U937), Human acute myelocytic leukemia (KG-1A) and Human Umbilical Vein Endothelial (HUVEC) cell lines. Methods: The effect of methanolic extract on the inhibition of cell proliferation and cytotoxic activity was evaluated by Dye exclusion and Micro culture tetrazolium test (MTT) cytotoxicity assay. Cell death ELISA was employed to quantify the nucleosome production result from nuclear DNA fragmentation during apoptosis and determined whether the mechanism involves induction of apoptosis or necrosis. Results: The present results demonstrated that methanolic extract at 50 to 800 μg/ml dose and time-dependently suppressed the proliferation of KG-1A, U937 and Raji cells by more than 80% (p<0.01), with ascending order of IC50 values in 24: KG-1A (214.377 μg/ml), U937 (229.312 μg/ml) and Raji (239.692 μg/ml) when compared with a chemotherapeutic anticancer drug, paclitaxel (Toxol), confirming the tumour-selective cytotoxicity. The crude extract however did not exert any significant cytotoxic effect on normal cell line HUVEC (IC50>800 Ag/ml). Nucleosome productions in KG-1A, Raji and U937 cells were significantly increased respectively upon the treatment of Salvia officinalis L. extract. Conclusion: The Salvia officinalis L. extract was found dose and time-dependently inhibits the proliferation of lymphoma and leukemic cells possibly via an apoptosis-dependent pathway.

Article Title : Mass-Production and Characterization of Anti-CD20 Monoclonal Antibody in Peritoneum of Balb/c Mice

Master Auther : Behzad Baradaran

Journal Title and Number : Advanced Pharmaceutical Bulletin. 2013; 3(1): 109-113.

Authers : Sineh sepehr K, Baradaran B, Majidi J, Abdolalizadeh J, Aghebati L, Zare Shahneh F

Publish : 2013

summary : Purpose: Monoclonal antibodies are important tools are used in basic research as well as, in diagnosis, imaging and treatment of immunodeficiency diseases, infections and cancers. The purpose of this study was to produce large scale of monoclonal antibody against CD20 in order to diagnostic application in leukemia and lymphomas disorders. Methods: Hybridoma cells that produce monoclonal antibody against human CD20 were administered into the peritoneum of the Balb/c mice which have previously been primed with 0.5 ml Pristane. After twelve days, approximately 7 ml ascetic fluid was harvested from the peritoneum of each mouse. Evaluation of mAb titration was assessed by ELISA method. In the present study, we describe a protocol for large scale production of MAbs. Results: We prepared monoclonal antibodies (mAbs) with high specificity and sensitivity against human CD20 by hybridoma method and characterized them by ELISA. The subclass of antibody was IgG2a and its light chain was kappa. Ascetic fluid was purified by Protein-A Sepharose affinity chromatography and the purified monoclonal antibody was conjugated with FITC and Immunofluorescence was done for confirming the specific binding. Conclusion: The conjugated monoclonal antibody could have application in diagnosis B-cell lymphomas, hairy cell leukemia, B-cell chronic lymphocytic leukemia, and melanoma cancer stem cells.

Article Title : Downstream characterization of anti-TNF-α single chain variable fragment antibodies

Master Auther : Omidi Y

Journal Title and Number : Human Antibodies. 2012; 21: 1–8.

Authers : Abdolalizadeh J, Nouri M, Majidi J, Baradaran B, Barzegari A, Omidi Y.

Publish : 2012

summary : Using phage display technology (PDT), we have recently isolated single-chain variable fragment (scFv) antibodies (Ab) against some pivotal molecular markers involved in malignancies and systemic inflammation including human tumor necrosis factor-alpha (TNF-α, GI:367465798). Downstream purification and characterization of scFv antibodies is a challenging issue, which may impose substantial impacts on quality of the final product(s). Of various purificationmethods, affinity chromatography has widely been used in the pharmaceutical grade downstream processing (PGDP) of proteins (e.g., monoclonal antibody (mAb) and scFvs). To pursue an optimized PGDP, in the current study, we have capitalized PDT for upstream selection of anti-TNF-α scFvs and protein A affinity chromatography (PAAC) for downstream extraction and purification of the scFvs from the crude medium secreted and periplasmic fractions of HB2151 cells. The versatility of the PDT selection was validated using SDS-PAGE electrophoresis, western blot, dot blot, ELISAand fluorescence microscopy. SDS-PAGEwestern blot analyses displayed a 17 kDa scFv with high purity (> 98%), while dot blot and ELISA analyses showed high specificity and binding affinity of the purified scFv antibody fragments toward human TNF-α at nM range. Fluorescence microscopy further confirmed detection of TNF-α in Raji B lymphoblast cells. Finally, based on our findings, we believe that these PDT selected and PAAC processed scFvs may be used as targeting and/or therapy agent for TNF-α mediated diseases. Further, to increase the production yield, downstream purification techniques need to be optimized for the large scale production of scFv antibodies.

Article Title : Cytotoxic Effects of Alcoholic Extract of Dorema Glabrum Seed on Cancerous Cells Viability

Master Auther : Amirkhiz MB

Journal Title and Number : Cytotoxic Effects of Alcoholic Extract of Dorema Glabrum Seed on Cancerous Cells Viability. Advanced Pharmaceutical Bulletin. 3(2): 403-408

Authers : Amirkhiz MB, Rashtchizadeh N, Nazemieh H, Abdolalizadeh J, Mohammadnejad L, Baradaran B

Publish : 2013

summary :

Article Title : Large Scale Generation and Characterization of Anti-Human CD34 Monoclonal Antibody in Ascetic Fluid of Balb/c Mice

Master Auther : Jafar Majidi

Journal Title and Number : Advanced Pharmaceutical Bulletin, 2013, 3(1), 211-216

Authers : Aghebati L, Majidi J, Baradaran B, Abdolalizadeh J, Kazemi T, Sineh sepehr K.

Publish : 2013

summary : Purpose: Monoclonal antibodies or specific antibodies are now an essential tool of biomedical research and are of great commercial and medical value. The purpose of this study was to produce large scale of monoclonal antibody against CD34 in order to diagnostic application in leukemia and purification of human hematopoietic stem/progenitor cells. Methods: For large scale production of monoclonal antibody, hybridoma cells that produce monoclonal antibody against human CD34 were injected into the peritoneum of the Balb/c mice which have previously been primed with 0.5 ml Pristane. 5 ml ascitic fluid was harvested from each mouse in two times. Evaluation of mAb titration was assessed by ELISA method. The ascitic fluid was examined for class and subclasses by ELISA mouse mAb isotyping Kit. mAb was purified from ascitic fluid by affinity chromatography on Protein A-Sepharose. Purity of monoclonal antibody was monitored by SDS -PAGE and the purified monoclonal antibody was conjugated with FITC. Results: Monoclonal antibodies with high specificity and sensitivity against human CD34 by hybridoma technology were prepared. The subclass of antibody was IgG1 and its light chain was kappa. Conclusion: The conjugated monoclonal antibody could be a useful tool for isolation, purification and characterization of human hematopoietic stem cells.

Article Title : Large Scale Generation and Characterization of Anti-Human CD34

Master Auther : Jafar Majidi1,Behzad Baradaran

Journal Title and Number : Advanced Pharmaceutical Bulletin, 2013, 3(1), 211-216

Authers : Leili Aghebati Maleki, Jafar Majidi, Behzad Baradaran, Jalal Abdolalizadeh, Tohid Kazemi, Ali Aghebati Maleki, Koushan Sineh sepehr

Publish : 2013

summary : Monoclonal antibodies or specific antibodies are now an essential tool of biomedical research and are of great commercial and medical value. The purpose of this study was to produce large scale of monoclonal antibody against CD34 in order to diagnostic application in leukemia and purification of human hematopoietic stem/progenitor cells. Methods: For large scale production of monoclonal antibody, hybridoma cells that produce monoclonal antibody against human CD34 were injected into the peritoneum of the Balb/c mice which have previously been primed with 0.5 ml Pristane. 5 ml ascitic fluid was harvested from each mouse in two times. Evaluation of mAb titration was assessed by ELISA method. The ascitic fluid was examined for class and subclasses by ELISA mouse mAb isotyping Kit. mAb was purified from ascitic fluid by affinity chromatography on Protein A-Sepharose. Purity of monoclonal antibody was monitored by SDS -PAGE and the purified monoclonal antibody was conjugated with FITC. Results: Monoclonal antibodies with high specificity and sensitivity against human CD34 by hybridoma technology were prepared. The subclass of antibody was IgG1 and its light chain was kappa. Conclusion: The conjugated monoclonal antibody could be a useful tool for isolation, purification and characterization of human hematopoietic stem cells.

Article Title : Investigating Apoptotic Effects of Methanolic Extract of Dorema glabrum Seed on WEHI-164 Cells.ISRN Pharmacology

Master Auther : Amirkhiz MB

Journal Title and Number : Investigating Apoptotic Effects of Methanolic Extract of Dorema glabrum Seed on WEHI-164 Cells.ISRN Pharmacology

Authers : Amirkhiz MB, Rashtchizadeh N, Nazemiyeh H, Abdolalizadeh J, Mohammadnejad L, Baradaran B

Publish : 2013

summary :

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