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Research field: recombinant proteins production ,purifications and characterization
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Research field: protein production and purification
Expert: Ebrahimi
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Address: TMU,faculty of medical sciences ,Building no 2
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Research field: phages in vaccines and anytbiotic therapy
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Dry eye syndrome (DES) and tear dysfunction are multifactorial conditions affecting meibomian glands, lacrimal glands, and ocular surface. This ocular disorder can cause eye irritation, irregular cornea, corneal barrier disruption, and blurred vision. Uncontrolled increase in matrix metalloproteinase-9 (MMP-9) level and activity has been detected in the tears and ocular surface in the patients with DES, which has been proved to be related to disruption of tight junctions in apical corneal epithelium associated with severe signs of DES. These uncontrolled activities of MMP-9 lead to desquamation of ocular surface epithelia. Therefore, this review study was conducted to summarize the evidence regarding MMP-9 contribution in DES, and inhibitio
PD-L1 overexpression on tumor cells forms a protective shield against cytotoxic T-cell killing, which consequently leads to immune evasion. Engagement of PD-1 in tumor infiltrating T cells with PD-L1 results in an exhausted T-cell phenotype, thus preventing an effective immune response against tumor cells. In the present study, we employed phage display combinatorial peptide library to discover anti-PD-L1 peptides. The peptides discovered here, could computationally exhibit specific interactions with PD-L1 at residues with which PD-1 also interacts. Binding affinity and specificity of the peptides were examined by flow cytometry. Anti- tumor activity of peptides was also investigated using several cell-based assays. Surprisingl
Lipid metabolism rewiring in gastric adenocarcinoma (GA) pathogenesis is still not clearly elucidated. This study aimed to describe the role of lipid catabolism in GA patient outcomes and possible therapeutic targets by analyzing the effect of hypoxia-inducible factor-1α (HIF-1α) on fatty acid oxidation (FAO). AGS cell line was cultured in normoxic and hypoxic conditions, and FAO-related genes were analyzed by real-time-PCR and Western-blot. The study group comprised 108 newly diagnosed GA patients and 152 control cases. Serum concentrations of medium and long-chain acyl-CoA dehydrogenases (MCAD and LCAD) proteins were measured using ELISA, and local expression of HIF-1α, carnitine palmitoyl transferase 1 (CPT1A) and peroxisome prolifera
The spike protein has been reported as one of the most critical targets for vaccine design strategies against the SARS-COV-2 infection. Hence, we have designed, produced, and evaluated the potential use of recombinant proteins derived from spike protein as vaccine candidates capable of neutralizing SARSCOV-2 virus.In silico tools were used to design spike-based subunit recombinant proteins (P1, P2, and P3). These proteins were checked for their ability to be identi ed by the anti-SARS-COV-2 antibodies by exposing them to Covid-19 serum samples. The proteins were then injected into mice and rabbits and the antibody titers were measured for 170 days. The virus neutralization test (VNT) was performed to analyze the obtained antibodies for thei
In this study, [111In]In‐DOTA‐PR81 was developed, and its preliminary preclinical qualifications were assessed for single photon emission computed tomography (SPECT) imaging of breast cancer. DOTA‐NHS‐ester was practiced and successively purified by molecular filtration. The chelate:mAb ratio was determined by spectrophotometry. DOTA‐PR81 was radiolabeled with In‐111 and its radiochemical yield, in vitro stability, in vitro internalization, and immunoreactivity tests were performed. SPECT imaging and tissue counting were applied to evaluate the tissue distribution of [111In]In‐DOTA‐hIgG and [111In]In‐DOTA‐PR81 in BALB/c mice bearing breast tumors. The radiochemical yield of [111In]In‐DOTA‐PR81 complex was >95.0 ? 0.5
Objective: The purpose of this study was to develop multivalent antibody constructs via grafting anti-HER2 antibodies, including Herceptin and oligoclonal-variable domain of heavy chain antibodies (VHHs), onto liposome membranes to enhance antibody activity and compare their effect on phospholipase C (PLC) signaling pathway with control.Materials and Methods: In this experimental study, SKBR3 and BT-474 cell lines as HER2 positive and MCF10A cell line as normal cell were screened with anti-HER2 antibodies, including constructs of multivalent liposomal antibody developed with Herceptin and anti-HER2 oligoclonal-VHHs. To confirm the accuracy of the study, immunofluorescent assay, migration assay and immuno-liposome binding ability to HER2 wer
The antibacterial properties of egg yolk antibodies have been known for many years. Enhanced antibiotic resistance has resulted in increased need for using these antibodies as an alternative. In the present study, generation, capsulation, and inhibition growth properties of IgY directed against Salmonella enterica subsp. enterica serovar Infantis (SI) were evaluated. White Leghorn layer hens were immunized using whole cell of inactivated SI. Salmonella Infantis–specific antibody activities in sera and egg yolk were determined by ELISA. A total of 480 one-day-old male “Cobb 500” chicks were randomly divided into 8 groups, with 6 replications of 10 birds kept for 21?D. All birds from 7 challenged groups were orally inoculated with 1?mL
Single stranded nucleic acids e.g. exosomal microRNAs have been utilized widely for analysis of pathological status of individuals, in recent years. Template enhanced hybridization process (TeHyP) is one of the promising strategies for detection and quantification of such nucleic acid biomarkers. In a TeHyP strategy two separate DNA strands only assemble for their performance when the target template is present. A TeHyP strategy may be combined with a split G-quadruplex peroxidase mimic to be a colorimetric assay of single stranded nucleic acids. In this study, a special case of such TeHyP was designed and investigated for its performance for a microRNA that was reported as a biomarker for triple negative breast cancer. The performance of t
The last day of December when China first reported to the World Health Organization (WHO) about unknown cause pneumonia detected in Wuhan, nobody was thinking about a pandemic of new corona virus throughout the world1. On 23 January 2020, the central government of China imposed a lockdown in Wuhan and other cities in Hubei in an effort to quarantine the outbreak of COVID-19; Although according to the declaration of WHO it was" unprecedented in public health history", it could fairly help them to control the disease2. On 11 February 2020, WHO announced the name of COVID-19 for the severe acute respiratory syndrome induced by a new coronavirus, SARS-CoV-23. It lasts only 70 days from the first report to the WHO until the official announcement
Glucose‐regulated protein 78 (GRP78) is an endoplasmic reticulum (ER) chaperone that has been shown that is overexpressed in cancer cells. Overexpression of GRP78 on cancer cells makes this molecule a suitable candidate for cancer detection and targeted therapy. VHH is the binding fragment of camelid heavy‐chain antibodies also known as “nanobody.” The aim of this study is to isolate and produce a new recombinant nanobody using phage display technique to detect cancer cells. Using the c‐terminal domain of GRP78 (CGRP) as an antigen, four rounds of biopanning were performed, and high‐affinity binders were selected by ELISA. Their affinity and functionality were characterized by surface plasmon resonance (SPR) cell ELISA and immu
Purpose To investigate the efficacy of RSH-12, a novel selective matrix metalloproteinase 9 (MMP-9) inhibitor peptide in rabbit models of dry eye syndrome (DES). Methods In vitro toxicity of RSH-12 on cultured human corneal fibroblasts was investigated with MTT. Ocular toxicity of RSH-12 was investigated by clinical examinations, histology, and TUNEL assay. Experimental model of dry eye was induced by 1.0% atropine sulfate administration followed after 15?min by treatment with PBS, RSH-12, and Restasis in individual groups, three times a day for 7?days. In addition to performing Schirmer’s test for evaluating basic tear secretion and tear b
The interaction between the angiotensin-converting enzyme 2 (ACE2) and the receptor binding domain (RBD) of the spike protein from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays a pivotal role in virus entry into the host cells. Since recombinant ACE2 protein has been suggested as an anti-SARS-CoV-2 therapeutic agent, this study was conducted to design an ACE2 protein with more desirable properties. In this regard, the amino acids with central roles in enzymatic activity of the ACE2 were substituted. Moreover, saturation mutagenesis at the interaction interface between the ACE2 and RBD was performed to increase their interaction affinity. The best mutations to increase the structural and thermal stability of the ACE2 wer
Calcium-sensing receptor (CaSR), which is better known for its action as regulating calcium homeostasis, can bind various ligands. To facilitate research on CaSR and understand the receptor's function further, an in silico designed truncated protein was developed. The resulting protein folding indicated that 99% of predicted three dimensional (3D) structure residues are located in favored and allowed Ramachandran plots. However, it was found that such protein does not fold properly when expressed in prokaryotic host cells. Thioredoxin (Trx) tag was conjugated to increase the final protein's solubility, which could help obtain the soluble antigen with better immunogenic properties. The truncated recombinant proteins were expressed and purifi
Methods: DOTA-NHS-ester was practiced and successively purified by molecular filtration. The chelate: mAb ratio was determined by spectrophotometry. DOTA-PR81 was radiolabeled with In-111 and its radiochemical yield, in vitro stability, in vitro internalization and immunoreactivity tests were performed. SPECT imaging and tissue counting were applied to evaluate the tissue distribution of [111 In] In-DOTA-hIgG and [111 In] In-DOTA-PR81 in BALB/c mice bearing breast tumors.Results: The radiochemical yield of [111 In] In-DOTA-PR81 complex was> 95.0?0.5%(ITLC) and the specific activity was 170?44 MBq/mg. Conjugation reaction resulted in the average number of chelators attached to a mAb (c/a) of 3.4?0.3: 1. The radioimmunoconjugate showed immuno
Matrix metalloproteinases (MMPs) play critical roles in a multiple number of autoimmunity diseases progression and metastasis of solid tumor. Gelatinases including MMP‐2 and MMP‐9 are extremely overexpressed in multiple pathological processes. MMP‐9 and MMP‐2 breakdown the extracellular matrix component gelatin very efficaciously. Therefore, designing and expansion of MMPs inhibitors can be an engrossing plan for therapeutic intermediacy. Anyway, a wide range of MMPs inhibitors face failure in several clinical trials. Due to sequence and structural conservation across the various MMPs, achieving specific and selective inhibitors is very demanding. In the current study, a phage‐displayed peptide library was screened using active h
The main challenge in the development of antibody is to select the appropriate antigen particularly when a truncated protein is used for immunization or as vaccine antigen. In previous studies, fragment selection was mainly based on epitopes and less often on the structure. Fewer studies have paid attention to the prediction of the truncated protein 3D structure and retained its similarity in the native and truncated proteins. Here we used in silico analysis to select two fragments of Pyruvate Kinase M2 (PKM2), as a tumor marker. One fragment, M-tPKM2, had a shorter sequence with one epitope although the predicted 3D structure was similar to the native PKM2. The other fragment, R-tPKM2, had a longer sequence and thus more epitopes, but had