Jentashapir Journal of Cellular and Molecular Biology
Background: Colon cancer is one of the most diagnosed cancers in the modern world. Chemotherapy i... more Background: Colon cancer is one of the most diagnosed cancers in the modern world. Chemotherapy is one of the most relevant treatments for colon cancer, but this method of treatment has severe and sometimes irreversible side effects. Therefore, alternative treatments with fewer side effects are highly considered. Natural and herbal medicine has been proven to be a great source for these alternative treatment methods. Objectives: This study aimed to investigate the anti-proliferative and cytotoxic effects of Ocimum basillicum (OB) leaf aqueous extract on human colon cancer cell lines LS174T and COLO205 and the expression of apoptotic genes. Methods: Methyl thiazol tetrazolium (MTT) assay was performed to assess the cytotoxic effects of our extract on the aforementioned cell lines. The changes in the expression of BAX and BAD were analyzed by qPCR following the treatment of cells with our extract. Results: The results of the MTT assay indicated that the extract had a cytotoxic effect ...
Phage display technology (PDT), a combinatorial screening approach, provides a molecular diversit... more Phage display technology (PDT), a combinatorial screening approach, provides a molecular diversity tool for creating libraries of peptides/proteins and discovery of new recombinant therapeutics. Expression of proteins such as monoclonal antibodies (mAbs) on the surface of filamentous phage can permit the selection of high affinity and specificity therapeutic mAbs against virtually any target antigen. Using a number of diverse selection platforms (e.g. solid phase, solution phase, whole cell and in vivo biopannings), phage antibody libraries (PALs) from the start point provides great potential for the isolation of functional mAb fragments with diagnostic and/or therapeutic purposes. Given the pivotal role of PDT in the discovery of novel therapeutic/diagnostic mAbs, in the current review, we provide an overview on PALs and discuss their impact in the advancement of engineered mAbs.
Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) ... more Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) fragments. Using PDT, target molecule-specific phage-Ab clones are enriched through the “biopanning” process. The individual specific binders are screened by the monoclonal scFv enzyme-linked immunosorbent assay (ELISA) that may associate with inevitable false-negative results. Thus, in this study, three strategies were investigated for optimization of the scFvs screening using Tomlinson I and J libraries, including (1) optimizing the expression of functional scFvs, (2) improving the sensitivity of ELISA, and (3) preparing different samples containing scFvs. The expression of all scFv Abs was significantly enhanced when scFv clones were cultivated in the terrific broth (TB) medium at the optimum temperature of 30 °C. The protein A–conjugated with horseradish peroxidase (HRP) was found to be a well-suited reagent for the detection of Ag-bound scFvs in comparison with either anti-c-myc Ab ...
Abstract According to the latest statistics, cancer is expected to account for nearly 1800000 new... more Abstract According to the latest statistics, cancer is expected to account for nearly 1800000 new cases and about 607000 deaths in the US in 2019. Approximately 40% of people with cancer survive. This means that life expectancy is a little more than 1 in 3. Therefore, it is essential to detect this malignancy in early stage and predict therapeutic responses in patients in order to enhance the efficiency of treatment, decrease side-effects and optimize the cure process. One of the best solutions for early cancer diagnosis and management is tracking tumor markers. Cancerous or noncancerous cells produce these substances in response to cancer or some specific conditions. Although, onco-biomarkers are generated in normal situations, their concentration is much higher in the presence of tumors. These materials which most of them are proteins can be spotted in body fluids such as blood, serum and urine. An increased level of an oncobiomarker may be associated with the presence of cancer, however, it is not a definite indicator. For that reason, a combination of biomarkers and other diagnostic tests are always preferred. Due to the high cost, low speed and intricate instrumental requirements of existing assays such as enzyme linked immuno-sorbent assay, fluorescence and radiometric immunoassay for measurement of biomarkers, there is an extreme need for more inexpensive, swift, simple and portable detection methods. In recent years, the popularity of electrochemical biosensors are increasing in diagnostic fields. Significant improvements in electrochemical techniques, biological instrumentation and nanotechnology have paved the way for the emergence of novel platforms for detection of tumor markers. Importance of measurement of biomarkers by electrochemical-based affinity biosensors for early cancer diagnosis and monitoring have been emphasized by numerous articles. The main purpose of the present review is to give researchers a general idea of the recent studies about early cancer detection by EC-based biosensing devices, being pursued globally.
Purpose: Production of functional recombinant antibody fragments in the periplasm of E. coli is a... more Purpose: Production of functional recombinant antibody fragments in the periplasm of E. coli is a prerequisite step to achieve sufficient reagent for preclinical studies. Thus, the cost-effective and lab-scale production of antibody fragments demands the optimization of culture conditions. Methods: The culture conditions such as temperature, optical density (OD600) at induction, induction time, and IPTG concentration were investigated to optimize the functional expression of a phage-derived scFv molecule using a design of experiment (DoE). Additionally, the effects of different culture media and osmolyte supplements on the expression yield of scFv were examined. Results: The developed 2FI regression model indicated the significant linear effect of the incubation temperature, the induction time, and the induction OD600 on the expression yield of functional scFv. Besides, the statistical analysis indicated that two significant interactions of the temperature/induction time and the tem...
Jentashapir Journal of Cellular and Molecular Biology
Background: Colon cancer is one of the most diagnosed cancers in the modern world. Chemotherapy i... more Background: Colon cancer is one of the most diagnosed cancers in the modern world. Chemotherapy is one of the most relevant treatments for colon cancer, but this method of treatment has severe and sometimes irreversible side effects. Therefore, alternative treatments with fewer side effects are highly considered. Natural and herbal medicine has been proven to be a great source for these alternative treatment methods. Objectives: This study aimed to investigate the anti-proliferative and cytotoxic effects of Ocimum basillicum (OB) leaf aqueous extract on human colon cancer cell lines LS174T and COLO205 and the expression of apoptotic genes. Methods: Methyl thiazol tetrazolium (MTT) assay was performed to assess the cytotoxic effects of our extract on the aforementioned cell lines. The changes in the expression of BAX and BAD were analyzed by qPCR following the treatment of cells with our extract. Results: The results of the MTT assay indicated that the extract had a cytotoxic effect ...
Phage display technology (PDT), a combinatorial screening approach, provides a molecular diversit... more Phage display technology (PDT), a combinatorial screening approach, provides a molecular diversity tool for creating libraries of peptides/proteins and discovery of new recombinant therapeutics. Expression of proteins such as monoclonal antibodies (mAbs) on the surface of filamentous phage can permit the selection of high affinity and specificity therapeutic mAbs against virtually any target antigen. Using a number of diverse selection platforms (e.g. solid phase, solution phase, whole cell and in vivo biopannings), phage antibody libraries (PALs) from the start point provides great potential for the isolation of functional mAb fragments with diagnostic and/or therapeutic purposes. Given the pivotal role of PDT in the discovery of novel therapeutic/diagnostic mAbs, in the current review, we provide an overview on PALs and discuss their impact in the advancement of engineered mAbs.
Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) ... more Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) fragments. Using PDT, target molecule-specific phage-Ab clones are enriched through the “biopanning” process. The individual specific binders are screened by the monoclonal scFv enzyme-linked immunosorbent assay (ELISA) that may associate with inevitable false-negative results. Thus, in this study, three strategies were investigated for optimization of the scFvs screening using Tomlinson I and J libraries, including (1) optimizing the expression of functional scFvs, (2) improving the sensitivity of ELISA, and (3) preparing different samples containing scFvs. The expression of all scFv Abs was significantly enhanced when scFv clones were cultivated in the terrific broth (TB) medium at the optimum temperature of 30 °C. The protein A–conjugated with horseradish peroxidase (HRP) was found to be a well-suited reagent for the detection of Ag-bound scFvs in comparison with either anti-c-myc Ab ...
Abstract According to the latest statistics, cancer is expected to account for nearly 1800000 new... more Abstract According to the latest statistics, cancer is expected to account for nearly 1800000 new cases and about 607000 deaths in the US in 2019. Approximately 40% of people with cancer survive. This means that life expectancy is a little more than 1 in 3. Therefore, it is essential to detect this malignancy in early stage and predict therapeutic responses in patients in order to enhance the efficiency of treatment, decrease side-effects and optimize the cure process. One of the best solutions for early cancer diagnosis and management is tracking tumor markers. Cancerous or noncancerous cells produce these substances in response to cancer or some specific conditions. Although, onco-biomarkers are generated in normal situations, their concentration is much higher in the presence of tumors. These materials which most of them are proteins can be spotted in body fluids such as blood, serum and urine. An increased level of an oncobiomarker may be associated with the presence of cancer, however, it is not a definite indicator. For that reason, a combination of biomarkers and other diagnostic tests are always preferred. Due to the high cost, low speed and intricate instrumental requirements of existing assays such as enzyme linked immuno-sorbent assay, fluorescence and radiometric immunoassay for measurement of biomarkers, there is an extreme need for more inexpensive, swift, simple and portable detection methods. In recent years, the popularity of electrochemical biosensors are increasing in diagnostic fields. Significant improvements in electrochemical techniques, biological instrumentation and nanotechnology have paved the way for the emergence of novel platforms for detection of tumor markers. Importance of measurement of biomarkers by electrochemical-based affinity biosensors for early cancer diagnosis and monitoring have been emphasized by numerous articles. The main purpose of the present review is to give researchers a general idea of the recent studies about early cancer detection by EC-based biosensing devices, being pursued globally.
Purpose: Production of functional recombinant antibody fragments in the periplasm of E. coli is a... more Purpose: Production of functional recombinant antibody fragments in the periplasm of E. coli is a prerequisite step to achieve sufficient reagent for preclinical studies. Thus, the cost-effective and lab-scale production of antibody fragments demands the optimization of culture conditions. Methods: The culture conditions such as temperature, optical density (OD600) at induction, induction time, and IPTG concentration were investigated to optimize the functional expression of a phage-derived scFv molecule using a design of experiment (DoE). Additionally, the effects of different culture media and osmolyte supplements on the expression yield of scFv were examined. Results: The developed 2FI regression model indicated the significant linear effect of the incubation temperature, the induction time, and the induction OD600 on the expression yield of functional scFv. Besides, the statistical analysis indicated that two significant interactions of the temperature/induction time and the tem...
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