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Osteoarthritis (OA) is the most common joint disease worldwide. Routine treatment options are limited, and total knee replacement surgeries often come with complications. In recent years, the use of biologics, such as Wharton’s jelly (Wj) derived from the umbilical cord (UC), has gained popularity. While mesenchymal stem cells (MSCs) derived from Wj show promise in restoring articular cartilage, they also have some limitations. Recent studies have indicated that exosomes isolated from acellular Wj may offer advantages under certain conditions.

To investigate the anti-inflammatory properties of exosomes isolated from Wj in synoviocytes.

Decellularization of Wj was performed using sterile umbilical cords obtained from patients. Next, the exosomes were isolated from Wj using ultracentrifugation. After characterizing the exosomes, they were co-cultured with inflammatory synovial fibroblast cells (HIG-82) for 24 hours. Then, the gene expression levels and protein contents of some important inflammatory mediators including metalloproteinase-13 (MMP-13), cyclooxygenase-2 (COX2) and inducible nitric oxide synthase (iNOS) were measured in the cells using real-time PCR and ELISA tests, respectively.

The expression levels of MMP-13, COX-2, and iNOS genes were significantly reduced in the cultured cells treated with exosomes compared to untreated cells. Moreover, the content of MMP-13, COX-2, and iNOS proteins were significantly lower in the supernatant of the cultured cells compared to the control.

Wj-derived exosomes exhibit notable anti-inflammatory properties, which can help mitigate inflammation in the synovial environment of joints. However, further research is required to fully understand their benefits and potential applications in treating osteoarthritis.

Recurrent pregnancy loss is defined as at least two consecutive clinical pregnancy losses before 20th week of gestation. In this study, we investigated microRNA-1 and microRNA-1229 in recurrent miscarriage patients before and after lymphocyte therapy in comparison to control group.

First, a written and informed consent will be obtained from the target population of this study, who are people with RPL referring to Waliasr International Hospital. According to the standard protocol, the number of 2×107 is injected subcutaneously to patients. Real-time PCR technique will be used to check the expression level of microRNA-1 and microRNA-1229.

The gene expression level of micro-RNA 1 in RPL women before of lymphocyte therapy was 2.030±1.445 and the gene expression level of the same micro-RNA after the lymphocyte therapy of the mother was 1.101±0.4780. Also, the expression level of micro-RNA 1229 in these women was reported to be 2.100±0.6296 before treatment and 1.247±0.9631 after treatment with lymphocytes. This level of gene expression in control group, for micro-RNA 1 and micro-RNA 1229 genes is 1.000±0.08334 and 1.000±0.08091, respectively.

The results of our study showed a noticeable decrease in the gene expression of microRNAs under study, both microRNAs 1 and 1229, in women with a history of three consecutive injections of the father's subcutaneous lymphocytes.

Recent studies show that most crops and horticultural plants can form symbiosis with the arbuscular mycorrhizal fungi (AMF) and the endophytic Serendepita indica, simultaneously. The endophytic fungus plays an important role in alleviating environmental stresses in plants. It has also been shown that excessive available phosphorus in soil limits the root colonization by arbuscular mycorrhizal fungi. No information is available on how soil phosphorus affects the establishment of endophytic fungus in root. Barley roots can be colonized by both mycorrhizal fungi and the endophytic fungus Serendipita indica. The objective of this study was to evaluate the effects of single or dual inoculation with Rhizophagus irregularis and Serendipita indica on barley roots under different phosphorus (P) levels. The researchers utilized a monoclonal antibody called MAb32B11 to assess the presence of glomalin, a signature molecule of arbuscular mycorrhizal (AM) fungi, in the roots. The glomalin content was quantified using the enzyme-linked immunosorbent assay (ELISA) method with the MAb32B11 antibody.

In this experiment, barley plants were inoculated with Rhizaphagus irregularis (AMF) and Serendepita indica (endophytic fungus) with three levels of phosphorus from triple super phosphate source. At the end of the vegetative growth period (about three months), the plants were harvested and phosphorus concentration in the plant were measured. A subsample from roots was stored in -20 ºC for determination of glomalin content. The glomalin content in the roots was analyzed using the monoclonal antibody MAb32B11. This antibody was employed to differentiate between the two fungi present in the roots and to quantify the abundance of arbuscular mycorrhizal fungi (AMF) specifically in plants treated with Rhizophagus irregularis. Additionally, the content of glomalin in the soil was determined at the end of the experiment using the same method as described above. The experiment was designed as a factorial completely randomized design (CRD) with three replications.

The results showed that the fresh and dry weights of shoot and root increased significantly in dual inoculation. At zero phosphorus level, shoot and root phosphorus concentrations were significantly higher in treatments with R. irregularis than in those without fungus (control). Under individual inoculation with R. irregularis, or S. indica as well as their dual inoculations, increasing level of phosphorus had no significant effect on shoot and root phosphorus concentration. In dual inoculation, the percentage of total colonization (88%) was significantly higher than that of individual inoculation treatment (68%) but the contribution of each fungus in root colonization under dual inoculation was significantly reduced as estimated by glomalin content of root and determination of total colonization. It was found that with increasing phosphorus level, total colonization percentage significantly decreased and the highest percentage of colonization (61%) was observed at zero level of phosphorus. By increasing phosphorus level, the percentage of root colonization was significantly decreased in individual inoculation by R. irregularis, or S. indica as well as dual inoculation. Results of glomalin assay in soil showed that the glomalin content was high in treatments of R. irregularis but control treatments without fungus and individual inoculation with S. indica had low glomalin. Antibody-reactive root glomalin was less in the dual inoculation treatment (1006.9 µg.g-1) than in the R. irregularis treatment alone (1924.5 µg.g-1) indicating that the presence of S. indica, in root inhibits, root colonization by R. irregularis. Moreover, the increasing of phosphorus level, significantly decreased root glomalin.

The increase of available phosphorus concentration in the soil caused to limit the expansion of the symbiosis of R. irregularis and S. indica, and this limitation was more for R. irregularis. In the case of dual inoculation with both Rhizophagus irregularis and Serendipita indica, the negative impact of phosphorus on colonization percentage was observed to be less compared to single inoculation. Although the percentage of colonization by each fungus decreased in the dual inoculation treatment compared to their individual inoculation, the overall colonization percentage increased significantly. It appears that in the dual inoculation scenario, while the total root colonization percentage increases, the presence of S. indica leads to a decrease in the colonization percentage specifically with R. irregularis. But in general, growth and nutrient absorption in the case of dual inoculation was better than the inoculation of each of them individually. It was also found that increasing the concentration of phosphorus in the soil caused a decrease in root colonization for both fungi, although the negative effect of phosphorus on the colonization of R. irregularis was more than that of S. indica. The measurement of glomalin in soil and root showed that the inhibitory effect of S. indica fungus on R. irregularis is less in soil than in root.

Sustainable agriculture is achievable by establishing a balance between plant and soil, and depends on the ability of soil and plant to support native and diverse microorganisms such as mycorrhizal fungi. These fungi by increasing the growth of the host plant and the development and stimulation of root secretions especially glomalin, plays an important role in considerable stability in soil ecosystem. Changing rangelands to agricultural uses can affect the symbiosis of these fungi and endanger the stability of ecosystems. This study was conducted in an area of 310 km2 in Sarab plain, The wheat, alfalfa, and potato fields were considered as agricultural uses of neighboring rangelands as control soils. From each land use, 30 samples were taken from the rhizosphere soil and roots of the plants and a total of 120 samples were taken. The percentage of mycorrhizal colonization and the amount of root glomalin and some soil properties were measured.

The root colonization was the highest in alfalfa compared to other land uses. Root glomalin was not statistically different between land uses. Soil available phosphorus had positive effect on root colonization at lower content (< 50 mg kg-1) while colonization percent showed a marked decrease above this level.

Colonization of perennial plants was more than annual plants and available phosphorus was the most important soil property that had an effect on fungal colonization of plant roots. However, no significant relationship was observed between contents of root glomalin and soil available phosphorus in different land uses.

Candida albicans is the most common cause of invasive candidiasis, but in recent years the incidence of infections caused by other species such as Candida Kruzei, Candida glabrata, Candida tropicalis, Candida parapsilosis and Candida lusitania has increased. In the last decade, the treatment methods for invasive candidiasis have changed completely, and a successful treatment depends on the timely start of treatment, the selection of an effective drug, and the lack of resistance of the fungus to that particular drug. On the other hand, the widespread use of immunosuppressive drugs as well as organ transplants has all caused widespread problems in the treatment of invasive candidiasis. Together, these observations highlight a rationale for the immediate development of new immunotherapy methods to enhance antifungal therapy in immunocompromised hosts. The past decade has seen great advances in our understanding of fungal immunobiology, leading to a number of new molecular and cellular immunotherapy methods for invasive fungal infections. Therefore, the aim of this study was to review the common and new antifungal drugs in the treatment of invasive candidiasis and to discuss the role of immunotherapy in better prevention and control of the disease.

Adjuvants are essential to potentiate the immune response to inoculated antigens and play a central role in vaccine development. Alum is generally used as a classic adjuvant, although it does not stimulate proper immunity, and some of the immunized subjects have low or no antibody response. Efforts have been continued to find more efficient adjuvants for better antibody responses. In the present study, the efficacy of three formulations of adjuvants, i.e. Cysteine p Guanine Oligodeoxynucleotide (CpG ODN), alum, and Freund, in the production of monoclonal anti Hepatitis B Surface Antigen (HBsAg) antibodies was investigated.

To immunize mice, regular hepatitis B vaccine containing recombinant HBsAg and alum was used with CpG ODN or Freund adjuvants, and splenocytes of hyperimmunized mice were fused with murine myeloma Sp2/0 cells. Positive hybridomas were selected by Enzyme-Linked Immunosorbent Assay (ELISA) using HBsAg as coating antigen followed by a limited dilution process.

The results showed that by using all three formulations of adjuvants, monoclonal antibody (mAb) specific to HBsAg was successfully generated. It was also found that the mice immunized with (HBsAg + Alum) + CpG had the highest concentration of antibody production in serum and hybridoma supernatants as well as positive clones. Based on these findings, the addition of CpG ODN also induced a higher antibody response compared with Complete Freund’s Adjuvant (CFA).

Results of this study showed that CpG and Freund adjuvants could be efficient partners for alum in the immunization period of the process of monoclonal antibody production.

 Food is the fundamental need of human life and has nutrients that support growth and be healthy. Gastrointestinal tract (GI) microbiota involves beneficial microorganisms that develop therapeutic effects and are characterize as probiotics. The studies on appropriate probiotic strains have led to the separation and description of specific metabolic byproducts of probiotics named postbiotics. 

 A literature review has been performing using PubMed, Medline, and Scopus databases using combinations of search terms “probiotic, postbiotic, immune system, bioactivity, cancer, health-promoting, food, and pharmaceutical industry”. All experimental studies were enrolling for this explanatory review. Articles that did not have a full text were excluded.

 The probiotics must maintain their survival against inappropriate growth conditions of the processing, storage, distribution, preparation, and digestion system so that they can play their most health effects. Conversely, probiotic metabolites have successfully overcome these unfavorable conditions and maybe a good alternative to probiotics. Due to their specific chemical structure, safe profile, long shelf-life, and the fact that they contain various signaling molecules, postbiotics may have anti-inflammatory, immunomodulatory, antihypertensive, inhibiting abnormal cell proliferation, and antioxidant activities.

 Postbiotics is mimic the fundamental and clinical role of probiotics, and due to their unique characteristics, can be applied in a delivery system (pharmaceutical/functional foods) to achieve health-promoting, prevention, and treatment purposes.

Umbilical cord blood hematopoietic stem cells (UCB-HSCs) are an attractive source for transplantation. The generation of megakaryocyte-committed cells could lead to shorten period of thrombocytopenia after HSCs transplantation. Platelet lysate (PL) unlike fetal bovine serum (FBS) can prevent immune problems as well as avert transmission of certain diseases to the recipient. In this study, the authors aimed to assess the effect of PL on UCB CD34+ cells expansion and megakaryocyte differentiation.

In this experimental study, PL prepared and the subsequent isolation of UCB CD34+ cells were done by magnetic cell sorting. The isolated cells were cultivated in Iscove’s Modified Dulbecco’s medium (IMDM) supplemented with PL or FBS. Cell expansion was evaluated using Trypan blue. Furthermore, Flow cytometry using monoclonal antibodies (CD41-FITC and CD42b-PE) and the expression of specific genes including GATA1, GATA2, FLI1, NFE2, and RUNX1 via real-time PCR were performed to evaluate the megakaryocyte differentiation.

The results showed that PL insignificantly enhanced UCB CD34+ cell expansion (32.83± 8.47 fold in FBS and 41.67± 10.31 fold in PL containing media). Besides, flow cytometry results showed that expression of CD41 was increased markedly (37.81± 4.78 fold in FBS and 45.78 ± 7.37 in PL containing media, P-value <0.05) but the elevation of CD42b (10.53 ± 2.13 and 13.20 ± 2.06 in FBS and PL containing media, respectively) was not significant (P-value = 0.051). The results of real-time PCR demonstrated a notable increase in GATA binding protein 1 (1.58, P-value <0.01), GATA binding protein 2 (2.45, P-value <0.001), RUNX family transcription factor 1 (1.60, P-value <0.01), Fli-1 proto-oncogene (1.87, P-value <0.001) in PL supplemented media, however, the increase of Nuclear Factor-Erythroid 2 gene expression was not significant in PL supplemented media (P-value = 0.11).

PL improved UCB CD34+ cells expansion and megakaryocyte differentiation compared to FBS.

Transfusion of red blood cells (RBCs) is a supportive and common treatment in surgical care, trauma, and anemia. However, in vivo production of RBC seems to be a suitable alternative for blood transfusions due to the limitation of blood resources, the possibility of disease transmission, immune reactions, and the presence of rare blood groups. Cell cultures require serum-free or culture media supplemented with highly expensive animal serum, which can transmit xenoviruses. Platelet lysate (PL) can be considered as a suitable alternative containing a high level of growth factors and a low production cost.

Three-step culture media supplemented with PL or fetal bovine serum (FBS) were used for proliferation and differentiation of CD34+ umbilical cord blood stem cells to erythrocytes in co-culture with bone marrow mesenchymal stem cells (BM-MSCs). The cells were cultivated for 15 days and cell proliferation and expansion were assessed using cell counts at different days. Erythroid differentiation genes, CD71 and glycophorin A expression levels were evaluated.

Maximum hematopoietic stem cells (HSCs) proliferation was observed on day 15 in PL-containing medium (99±17×103-fold). Gene expression and surface markers showed higher differentiation of cells in PL-containing medium.

The results of this study indicate that PL can enhance erythroid proliferation and differentiation of CD34+ HSCs. PL can also be used as a proper alternative for FBS in the culture medium and HSCs differentiation.

As many investigations have reported, there is a complicated relation between fermented foods, lactic acid bacteria (LAB), and human health. It seems that bioactive components such as prebiotics, probiotics, and postbiotics are key mediators of the complex and direct association between these factors. LAB activity in the matrix of fermented foods and improving their growth by prebiotic compounds ultimately results in the production of bioactive molecules (postbiotics), which possess specific biological and physiological properties. The term "postbiotics" refers to a complex of biological micro- and macromolecules, if consumed in adequate amounts, provides the host with different health-promoting effects. Different reports have suggested that postbiotics possess the ability to moderate the effectiveness of cancer treatment and reduce the side-effects of conventional therapies in cancer patients due to their anti-proliferative, anti-inflammatory and anti-cancer properties. Consequently, postbiotics, for their unique characteristics, have gained great scientific attention and are considered as a novel approach for adjuvant therapy in patients with cancer.

Tumor microenvironment consists of malignant and non-malignant cells. The interaction of these dynamic and different cells is responsible for tumor progression at different levels. The non-malignant cells in TME contain cells such as tumor-associated macrophages (TAMs), cancer associated fibroblasts, pericytes, adipocytes, T cells, B cells, myeloid-derived suppressor cells (MDSCs), tumor-associated neutrophils (TANs), dendritic cells (DCs) and Vascular endothelial cells. TAMs are abundant in most human and murine cancers and their presence are associated with poor prognosis. The major event in tumor microenvironment is macrophage polarization into tumor-suppressive M1 or tumor-promoting M2 types. Although much evidence suggests that TAMS are primarily M2-like macrophages, the mechanism responsible for polarization into M1 and M2 macrophages remain unclear. TAM contributes cancer cell motility, invasion, metastases and angiogenesis. The relationship between TAM and tumor cells lead to used them as a diagnostic marker, therapeutic target and prognosis of cancer. This review presents the origin, polarization, role of TAMs in inflammation, metastasis, immune evasion and angiogenesis as well as they can be used as therapeutic target in variety of cancer cells. It is obvious that additional substantial and preclinical research is needed to support the effectiveness and applicability of this new and promising strategy for cancer treatment.

Fasting is one of the most basic religions in Islam. The body considers fasting as a stress and tries to cope with it through a path called self-regulating stress. The effects of fasting on various diseases and different physiological conditions have been evaluated and its positive effects have been shown in many diseases. The effects of fasting on the immune system of healthy people in general indicate the effect of fasting moderator on the chemokine network, oxidative stress and intrinsic and acquired immune system. Correspondingly, fasting reduces the pre-inflammatory cytokines, the chemokines, the antibodies, the CRP and the immune system cells, and increases IFN-γ, TNFα, iNOS, Endorphine and endocannabinoids. In addition, fasting does not have an effect on oxidative stress markers such as Malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase, and catalase levels. Fasting also has beneficial effects on autoimmune diseases and, in most cases, moderates the disease. Various studies have shown that fasting with chemotherapy can reduce the adverse effects of chemotherapy without even reducing the anti-cancer effect of chemotherapy drugs, and even reduce the growth of some cancers in vitro. Although, chemotherapy with fasting is a better option for cancer treatment for prognosis, there are very precise clinical trials prior to using it as a standardized method. In this review article, we discuss the significance of fasting effects on various diseases and different physiological conditions and their physiological elements on the chemokine network, oxidative stress and intrinsic immune system in cancer development.

Colorectal cancer (CRC) remains a universal and lethal cancer owing to metastatic and relapsing disease. Currently, the role of microRNAs has been checked in tumorigeneses. Numerous studies have revealed that between the tumor suppressor miRNAs, the reduced expression of miR-146a-5p and -193a-5p in several cancers including CRC tissues are related with tumor progression and poor prognosis of patients. The purpose of this study is to examine the role of miR-146 a-5p and -193 a-5p in CRC cell cycle progression.

The miR-193a-5p and -146 a-5p mimics were transfected into HT-29 CRC cells via jetPEI transfection reagent and their impact was assessed on p53, cyclin B, and NF-kB gene expression. The inhibitory effect of these miRNAs on cell cycle was assessed by flow cytometry. The consequence of miR-193a-5p and miR-146 a-5p on the protein expression level of Murine double minute 2 (MDM2) was assessed by western blotting.

miR193a-5p and -146a-5p regulated the expression of MDM2 protein and p53, cyclin B, and NF-kB gene expression in CRC cells. Treatment of HT-29 cells with miRNA-146a-5p and -193a-5p induced G1 cell cycle arrest.

The findings of our study suggest that miR146a-5p and -193a-5p may act as a potential tumor suppressor by their influence on cell cycle progression in CRC cells. Thus, miRNA-146a-5p and -193a-5p restoration may be recommended as a potential therapeutic goal in the treatment of CRC patients.

Although it has been frequently confirmed that HLA-G plays an important role in the reproduction and pregnancy, the pattern of HLA-G gene and its protein expression are rarely addressed in studies. Therefore we conducted this study in regard to evaluate the HLA-G gene and its protein expression in the women’s placenta with recurrent miscarriage. Placental samples were obtained from the women who were admitted for delivery or abortion in Al Zahra and Taleghani hospitals, Tabriz, Iran. HLA-G gene expression was determined by real-time polymerase chain reaction (PCR) and HLA-G protein expression was assessed by western blotting and immunofluorescence staining in the tissue samples. The results showed a significant decrease in the expression of gene and proteins of HLA-G in the women with recurrent miscarriage compared to the control placental tissues. According to the obtained results, it was concluded that the decrement of HLA-G gene and protein expressions are associated with recurrent miscarriage. Since there are conflicting results from other studies, it is suggested to conduct a more comprehensive similar study with greater sample size.

Glomalin is a specific glycoprotein produced by the fungi belonging to phylum Glomeromycota and plays a key role in soil carbon and nitrogen storage. This also has a significant role in the stable aggregates formation and establishment of microbial communities in soil. Assimilated plant C which is allocated to the mycorrhizal fungus, appears as a recalcitrant glycoprotein (glomalin) in cell walls of hyphae and spores. Considering global warming due to increasing greenhouse gases, this phenomenon cab be important in carbon sequestration and reducing CO2 in atmosphere. Chemical fertilizers can affect symbiotic relations of these fungi, which in turn affect glomalin production.
In a factorial completely randomized design with three replication, clover plants (Trifolium repense L.) were included with Rhizophagus irregularis and/or Rhizobium leguminosarum bv. Trifolii. Four levels of nitrogen (0, 2, 6 and 10 mM as nitrate) in Newman & Romheld nutrient solution were applied to the pots containing 1.5 kg sterile sand. The pots were daily irrigated with nutrient solution containing the above-mentioned levels of nitrogen. Clover plants were excised after 12 weeks of growth. Fine roots were cleaned with %10 KOH and then stained using lactoglycerol trypan blue. Root colonization percentage was determined by grid line intersections method (GLM) described by Norrif et al (1992). For glomalin extraction, hyphal or root samples were autoclaved at 121 ⁰C with 50 mM sodium citrate buffer for 60 min in three cycles. Sand glomalin (SG) and root glomalin (RG) were measured by Bradford method after extraction. Nitrogen concentration in shoot and root was measured according to the standard method.
By increasing nitrogen level, the SG significantly decreased (p Carbon sequestration via glomali synthase by AM fungi is an important pathway for capturing CO2 from atmosphere. Field management measures help AM development of glomalin production. Based on our results, co-inoculated plants with AM and rhizobuim seem to positively affect the production of this glycoprotein. On the other hand, SG decreased significantly by increasing nitrogen concentrations in the nutrient solution. RG, however, increased significantly as a result of increased nitrogen in both fungal inoculations. The highest amount of RG was recorded in the co-inoculated plants with 10mM level. Glomalin synthesis by the fungi is positively affected by the soil nitrogen availability. Nitrogen is the main constituent of this glycoprotein. Plant photosynthates are translocated to the fungal organs via roots and mainly utilized for glomalin synthesis in hyphal and spore cell walls. During this process, nitrogen plays an important role as a constituent of the glycoprotein. The Bradford method was used for glomalin determination in this study. The method is not specific for glomalin and can also measure other glomalin related proteins and glycoproteins. Other proteins increased by N fertilization can hence be measured based on Bradford method. Once plant assimilates are translocated to the fungi, they may be transformed to the nitrogenous compounds if sufficient nitrogen sources are available. Accordingly, a considerable amount of fixed carbon is assimilated in fungal organs and soil particles. It can be concluded that carbon sequestration by arbuscular mycorrhizal symbiosis in terrestrial ecosystems can be improved by N fertilization at optimum level. In addition, the presence of rhizobium bacteria can meet the nitrogen requirement of plants through biological stabilization of nitrogen.

Purified mouse IgG2a (a product that could be used in medical research) subclass could be used for animal immunization to production of polyclonal Antibody and to obtain hybridomas in Monoclonal Antibody Production Procedures. The goal of this study was to purify the mouse IgG2a.
In one step, Ion exchange chromatography was carried out for purification of mouse IgG and then in second step, ProA affinity chromatography was used for IgG2a purification .The chosen method for determination of purity was reduced and non-reduced SDS-PAGE. ELISA method was used for titer and isotype determination.
Mouse Igs with a protein concentration of 27mg/ml (volume: 3CC) was applied on Ion exchange column. Purification by Ion exchange chromatography yielded about 28mg of mouse IgG. Eight mg mouse IgG2a was obtained by ProA affinity chromatography. In reduced SDS-PAGE analysis of purified antibody, two bands were seen in 25& 50 KDa MW positions. Isotype determination of purified mouse IgG2a with mouse isotyping Kit showed the presence of mouse IgG2a isotype with a kappa light chain in related fraction.
Purified mouse IgG2a subclass was obtained with purity more than 95%. Due to the obtained high purity we concluded that Ion exchange chromatography following by ProA affinity chromatography could be a suitable method for purification of mouse IgG subclasses with high quality. Our product is an economical and suitable product that takes a step towards self-sufficiency of the country.

Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies are produced against different epitopes of antigens. Purified F(ab')2 can be used for animal’s immunization to produce polyclonal antibodies. Human immunoglobulin G (IgG) was purified by ion exchange chromatography method. In all stages verification method of the purified antibodies was sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purified IgG was digested by pepsin enzyme and F(ab')2 fragment was purified by gel filtration separation method. For production of polyclonal antibody, rabbit was immunized by purified F(ab')2 and antibody production was investigated by enzyme-linked immunosorbent assay. Purified anti-IgG F(ab')2 was conjugated with fluorescein isothiocyanate. Ion exchange chromatography purification yielded 38 mg of human IgG antibody. The results of SDS-PAGE in reduced and non-reduced conditions showed bands with 25-30 kDa molecular weight (MW) and 50-kDa respectively and a distinct band with 150 kDa MW. The results of non-reduced SDS-PAGE for determining the purity of F(ab')2 fragment showed one band in 90 kDa and a band in 150 kDa MW position. Purification by Ion exchange chromatography method resulted about 12 mg rabbit polyclonal antibody. Flow cytometry showed generated polyclonal antibody had an acceptable activity compared to commercial antibody. Taking together, purified IgG F(ab')2 and polyclonal anti-IgG F(ab')2 are useful tools in biomedical and biochemical researches and diagnostic kits.

Surface plasmon resonance (SPR) sensing confers a real-time assessment of molecular interactions between biomolecules and their ligands. This approach is highly sensitive and reproducible and could be employed to confirm the successful binding of drugs to cell surface targets. The specific affinity of monoclonal antibodies (MAb) for their target antigens is being utilized for development of immuno-sensors and therapeutic agents. CD20 is a surface protein of B lymphocytes which has been widely employed for immunotargeting of B-cell related disorders. In the present study, binding ability of an anti-CD20 MAb to surface antigens of intact target cells was investigated by SPR technique.
Two distinct strategies were used for immobilization of the anti-CD20 MAb onto gold (Au) chips. MUA (11-mercaptoundecanoic acid) and Staphylococcus aureus protein A (SpA) were the two systems used for this purpose. A suspension of CD20-positive Raji cells was injected in the analyte phase and the resulting interactions were analyzed and compared to those of MOLT-4 cell line as CD20-negative control.
Efficient binding of anti-CD20 MAb to the surface antigens of Raji cell line was confirmed by both immobilizing methods, whereas this MAb had not a noticeable affinity to the MOLT-4 cells.
According to the outcomes, the investigated MAb had acceptable affinity and specificity to the target antigens on the cell surface and could be utilized for immunodetection of CD20-positive intact cells by SPR method.

The ability of polyclonal antibodies to react with many epitopes of an antigen makes them valuable reagents in research and diagnosis. The aim of this study was purification of mouse IgG2a and production of polyclonal antibody against purified mouse IgG2a subclass.
Mouse IgG2a was purified by ProA affinity. Verification method of the purified antibody was SDS-PAGE and ELISA by a mouse isotyping Kit. Rabbit was immunized with purified IgG2a. The production of antibody in rabbit was investigated by direct ELISA method. Rabbit serum was collected and precipitated at the final concentration of 50% ammonium sulfate. Polyclonal antibody was purified by ion-exchange chromatography and labeled with HRP. The titre and cross reactivity of product was detected by direct ELISA method.
The results of SDS-PAGE in reduced and non-reduced conditions showed bands with 50-KDa, 25-30 KDa MW and a distinct band with 150 KDa MW. Isotype determination showed the presence of mouse IgG2a in related fraction. The titer of Anti-mouse polyclonal antibody was 200000. The optimum titer of prepared HRP conjugated IgG was 4000. Conjugated rabbit IgG has more cross reactivity with mouse IgG2b.
Taking together, affinity chromatography and ion-exchange chromatography are appropriate techniques for purification of mouse IgG subclasses and rabbit IgG, respectively.

Mouse IgG subclasses containing IgG1, IgG2a, IgG2b and IgG3 have been defined and described both physiochemically and immunologically. Sepharose beads conjugated with protein A affinity chromatography was used for purification of mouse IgG2b. Sodium citrate buffer (0.1 M, pH: 3.5) was used for separation of mouse IgG2b. Verification of the purified fractions was monitored by SDS-PAGE (polyacrylamide gel electrophoresis) in reducing condition. Immunized rabbit serum was collected and precipitated at the final concentration of 50% ammonium sulfate. After dialysis against tris-phosphate buffer (pH: 8.1) ion exchange chromatography column was used for purification of rabbit anti-mouse IgG2b. The periodate method was performed for conjugation with some variations. After conjugation, direct ELISA was used to determine the titer of HRP conjugated rabbit IgG against mouse IgG2b. The titer of rabbit anti-mouse IgG2b that determined by ELISA was 32000. The purity of rabbit anti-mouse IgG2b was about 95%. The optimum dilution of prepared HRP conjugated IgG was 1:10000. This study showed that ion-exchange chromatography and affinity chromatography could be appropriate techniques for purification of mouse IgG and IgG subclasses respectively. This study showed that affinity chromatography could be an appropriate method for purification of IgG2b antibodies.

Despite significant advances have been achieved in cancer therapy, response to conventional treatments like surgery, radiotherapy and chemotherapy varies among individuals. Immunotherapy is known to be an effective strategy for patients who are resistant to the currently available interventions. Ninety-six male Balb/c mice (aged 6-8 weeks) were selected and divided into twelve groups of eight. Approximately, 1×106of WEHI-164 cells were injected to each mouse for tumor genesis. Five immunotherapy treatments were considered in this study, including Heat Shock Proteins (HSP), Bacillus Calmette-Guérin (BCG), Bifidobacterium, Immuno-Modulator Drug (IMOD) and Thalidomide. After tumor formation, the groups were treated with one or more of these therapies. Tumor size and survival rate was regularly monitored. Depending on the treatment group, tumor sizes were different. In some groups, combined treatments demonstrated more inhibitory effects on tumor growth rate. The mice in group (IMOD+ Thalidomide) had the lowest survival rate but group (BCG+ HSP+ Thalidomide) survived until the end of the experiment. The (HSP+ BCG+ Thalidomide) group exhibited satisfactory outcomes and two third of the mice in this group went into complete remission. Some combination therapies in test groups had significant impacts on survival and tumor growth rate.

CD34 is a type I membrane protein with a molecular mass of approximately 110 kDa. This antigen is associated with human hematopoietic progenitor cells and is a differentiation stage-specific leukocyte antigen. In this study we have generated and characterized monoclonal antibodies (mAbs) directed against a CD34 marker. Mice were immunized with two keyhole lympet hemocyanin (KLH)-conjugated CD34 peptides. Fused cells were grown in hypoxanthine, aminopterine and thymidine (HAT) selective medium and cloned by the limiting dilution (L.D) method. Several monoclones were isolated by three rounds of limited dilutions. From these, we chose stable clones that presented sustained antibody production for subsequent characterization. Antibodies were tested for their reactivity and specificity to recognize the CD34 peptides and further screened by enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses. One of the mAbs (3D5) was strongly reactive against the CD34 peptide and with native CD34 from human umbilical cord blood cells (UCB) in ELISA and Western blotting analyses. The results have shown that this antibody is highly specific and functional in biomedical applications such as ELISA and Western blot assays. This monoclonalantibodies(mAb) can be a useful tool for isolation and purification of human hematopoietic stem cells (HSCs).

Monoclonal antibodies and related conjugates are key reagents used in biomedical researches as well as, in treatment, purification and diagnosis of infectious and non- infectious diseases. Balb/c mice were immunized with purified human IgG. Spleen cells of the most immune mouse were fused with SP2/0 in the presence of Poly Ethylene Glycol (PEG). Supernatant of hybridoma cells was screened for detection of antibody by ELISA. Then, the sample was assessed for cross-reactivity with IgM & IgA by ELISA and confirmed by immunoblotting. The subclasses of the selected mAbs were determined. The best clone was injected intraperitoneally to some pristane-injected mice. Anti-IgG mAb was purified from the animal's ascitic fluid by Ion exchange chromatography and then, mAb was conjugated with HRP. In the present study, over than 50 clones were obtained that 1 clone had optical density over than 3. We named this clone as supermonoclone which was selected for limiting dilution. The result of the immunoblotting, showed sharp band in IgG position and did not show any band in IgM&IgA position. Based on the findings of this study, the conjugated monoclonal antibody could have application in diagnosis of infectious diseases like Toxoplasmosis, Rubella and IgG class of other infectious and non- infectious diseases.

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. 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. 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. The conjugated monoclonal antibody could be a useful tool for isolation, purification and characterization of human hematopoietic stem cells.