ABSTRACT
Cancer stem cells (CSCs) with their self-renewal ability are accepted as cells which initiate tumors. CSCs are regarded as interesting targets for novel anticancer therapeutic agents because of their association with tumor recurrence and resistance to conventional therapies, including radiotherapy and chemotherapy. Chimeric antigen receptor (CAR)-T cells are engineered T cells which express an artificial receptor specific for tumor associated antigens (TAAs) by which they accurately target and kill cancer cells. In recent years, CAR-T cell therapy has shown more efficiency in cancer treatment, particularly regarding blood cancers. The expression of specific markers such as TAAs on CSCs in varied cancer types makes them as potent tools for CAR-T cell therapy. Here we review the CSC markers that have been previously targeted with CAR-T cells, as well as the CSC markers that may be used as possible targets for CAR-T cell therapy in the future. Furthermore, we will detail the most important obstacles against CAR-T cell therapy and suggest solutions.
ABSTRACT
Background: 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
Methods: 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
Results: 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 25and 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
Conclusion: 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
ABSTRACT
Republic of Azerbaijan is considered as an area with high prevalence of multidrug resistant tuberculosis. Uncontrolled travelling of Azerbaijanis people to Iran is the issue that needs to be considered as an important issue. This study was conducted on 32 patients with tuberculosis from Baku-Nakhchivan and 48 patients from Iran during 2012 to 2014. Colonies of Mycobacterium tuberculosis were examined after isolating them from patients using proportional method on Lowenstein-Jensen media regarding resistance encounter with Rifampin, Isoniazid and Ethambutol. Among M. tuberculosis isolates belonging to 32 foreign patients; 69%, 72% and 56% of them were resistant to Rifampin, Isoniazid and Ethambutol, respectively [multidrug resistance tuberculosis: MDR-TB: 62.5%]. From 48 isolates of Iranian patients; 8%, 4% and 4% were resistant to Rifampin, Isoniazid and Ethambutol, respectively [MDR-TB: 2.1%]. Resistant strains are common in people from Baku-Nakhchivan. To prevent the transmission of these strains to Iranians, strategies such as; establishing a medical campus in border lines of both countries for clinical examinations and conducting screening tests regarding tuberculosis infection in applicants for entering Iran must be taken in to account
ABSTRACT
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 monoclonal antibodies [mAb] can be a useful tool for isolation and purification of human hematopoietic stem cells [HSCs]