Breast cancer is the most common cancer among women in the world. One of the approaches for diagnosis of breast cancer is detection of its tumor-associated markers. Mucin 1 (MUC1), a tumor-associated antigen, is a transmembrane glycoprotein expressed by normal epithelial cells and overexpressed by carcinomas of epithelial origin. Also, human epidermal growth factor receptor-2 (HER2/erbB-2) belongs to the one of four members of tyrosin kinase type 1 family in which overexpression of HER2 is associated with malignancy in breast cancer. This study was aimed to bioinformatics analysis and designing a recombinant chimeric protein containing MUC1 and HER2 antigens to express in prokaryotic host (Escherichia coli) as breast cancer diagnosis tools. The immunogenic sequences of MUC1 and HER2 were extracted and fused together by a linker. The chimeric construct was analyzed by bioinformatics softwares. The optimization and purification, evaluation of the expression of chimeric protein was performed using Western blotting, ELISA, and immunohistochemistry. The results showed that the chimeric construct was stable and immunogenic domains were exposed. The pET-28a vector containing chimeric gene had high level of protein expression. The recombinant chimeric protein was confirmed using Western blotting, and it was investigated using ELISA and IHC. Then, the MUC1 and HER2 combined peptides can be used as coating antigens in ELISA for detection of antibodies against MUC1 or HER2 in human serum.
High desalination performance, dye retention, and antibacterial properties were achieved with a multifunctional thin-film nanocomposite (MTFN) membrane, fabricated by the incorporation of a novel nanocomposite structure of reduced graphene oxide@TiO2@Ag (rGO@TiO2@Ag) into the polyamide active layer. The specific characteristics of the graphene-based nanocomposite, synthesized by the microwave-assisted irradiation process, favored water channelization and provided superhydrophilicity and antibacterial properties to the MTFN membranes. In comparison with the conventional methods, such as multistep chemical process using strong agents for reduction and long-term energy-consuming hydrothermal process, microwave irradiation facilitated a green, fast, and cost-effective route for the fabrication of GO-based nanocomposites for multifunctional applications. Interfacial polymerization was performed on a polyethersulfone/Si3N4 robust hollow fiber substrate using m-phenylenediamine aqueous solution and 1,3,5-benzenetricarbonyltrichloride organic solution. The structural and chemical characteristics of the synthesized nanocomposites and the MTFN membranes were thoroughly studied by a series of characterization analyses (transmission electron microscopy, field emission scanning electron microscopy, X-ray diffraction, Fourier transform infrared, energy-dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy, and atomic force microscopy). The physicochemical properties and the nanofiltration performance of the MTFN membranes were investigated after the incorporation of rGO@TiO2@Ag at various concentrations. The water contact angles confirmed the superb surface hydrophilicity of the MTFN membranes. High permeability (52 L·m–2·h–1), desalination (96% for 1 g/L Na2SO4 feed solution), and dye retention (98% for 0.5 g/L rose bengal feed solution) were recorded for MTFN enriched with 0.2 wt % rGO@TiO2@Ag. A 90% reduction in the number of viable bacteria (Escherichia coli), after 3 h of contact with MTFN membranes, confirmed the superior antibacterial activity of the produced membranes.
Recombinant Escherichia coli displaying organophosphorus hydrolase (OPH) was used to overcome the diffusion barrier limitation of organophosphorus pesticides. A new anchor system derived from the N-terminal domain of ice-nucleation protein from Pseudomonas syringae InaV (InaV-N) was used to display OPH onto the surface. The designed sequence was cloned in the vector pET-28a(+) and then was expressed in E. coli. Tracing of the expression location of the recombinant protein using SDS-PAGE showed the presentation of OPH by InaV-N on the outer membrane, and the ability of recombinant E. coli to utilize diazinon as the sole source of energy, without growth inhibition, indicated its significant activity. The location of OPH was detected by comparing the activity of the outer membrane fraction with the inner membrane and cytoplasm fractions. Studies revealed that recombinant E. coli can degrade 50% of 2 mM chlorpyrifos in 2 min. It can be concluded that InaV-N can be used efficiently to display foreign functional protein, and these results highlight the high potential of an engineered bacterium to be used in bioremediation of pesticide-contaminated sources in the environment.
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