Agar-tragacanth/silk fibroin hydrogel containing Zn-based MOF as a novel nanobiocomposite with biological activity.

In this study, a novel nanobiocomposite consisting of agar (Ag), tragacanth gum (TG), silk fibroin (SF), and MOF-5 was synthesized and extensively investigated by various analytical techniques and basic biological assays for potential biomedical applications. The performed Trypan blue dye exclusion assay indicated that the proliferation percentage of HEK293T cells was 71.19%, while the proliferation of cancer cells (K-562 and MCF-7) was significantly lower, at 10.74% and 3.33%. Furthermore, the Ag-TG hydrogel/SF/MOF-5 nanobiocomposite exhibited significant antimicrobial activity against both E. coli and S. aureus strains, with growth inhibition rates of 76.08% and 69.19% respectively. Additionally, the hemolytic index of fabricated nanobiocomposite was found approximately 19%. These findings suggest that the nanobiocomposite exhibits significant potential for application in cancer therapy and wound healing.

Chemical and physical modification of graphene oxide nano-sheets using casein, Zn-Al layered double hydroxide, alginate hydrogel, and magnetic nanoparticles for biomedical applications.

In our study, we developed a novel nanobiocomposite using graphene oxide (GO), casein (Cas), ZnAl layered double hydroxide (LDH), sodium alginate (Alg), and Fe3O4 magnetic nanoparticles. To synthesize the GO, we used a modified Hummer's method and then covalently functionalized its surface with Cas protein. The functionalized GO was combined with as-synthesized ZnAl LDH, and the composite was conjugated with alginate hydrogel through the gelation process. Finally, we magnetized the nanobiocomposite using in-situ magnetization. The nanobiocomposite was comprehensively characterized using FT-IR, FE-SEM, EDX, and XRD. Its biological potential was assessed through cell viability, hemolysis, and anti-biofilm assays, as well as its application in hyperthermia. The MTT assay showed high cell viability percentages for Hu02 cells after 24, 48, and 72 h of incubation. The nanobiocomposite had a hemolytic effect lower than 3.84 %, and the measured bacterial growth inhibition percentages of E. coli and S. aureus bacteria in the presence of the nanobiocomposite were 52.18 % and 55.72 %, respectively. At a concentration of 1 mg.mL-1 and a frequency of 400 kHz, the nanocomposite exhibits a remarkable specific absorption rate (SAR) of 67.04 W.g-1, showcasing its promising prospects in hyperthermia applications.

A novel ternary magnetic nanobiocomposite based on tragacanth-silk fibroin hydrogel for hyperthermia and biological properties.

This study involves the development of a new nanocomposite material for use in biological applications. The nanocomposite was based on tragacanth hydrogel (TG), which was formed through cross-linking of Ca2+ ions with TG polymer chains. The utilization of TG hydrogel and silk fibroin as natural compounds has enhanced the biocompatibility, biodegradability, adhesion, and cell growth properties of the nanobiocomposite. This advancement makes the nanobiocomposite suitable for various biological applications, including drug delivery, wound healing, and tissue engineering. Additionally, Fe3O4 magnetic nanoparticles were synthesized in situ within the nanocomposite to enhance its hyperthermia efficiency. The presence of hydrophilic groups in all components of the nanobiocomposite allowed for good dispersion in water, which is an important factor in increasing the effectiveness of hyperthermia cancer therapy. Hemolysis and 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays were conducted to evaluate the safety and efficacy of the nanobiocomposite for in-vivo applications. Results showed that even at high concentrations, the nanobiocomposite had minimal hemolytic effects. Finally, the hyperthermia application of the hybrid scaffold was evaluated, with a maximum SAR value of 41.2 W/g measured in the first interval.

Fabrication and biological investigation of a novel star polymer based on magnetic cyclic aromatic polyimide chains.

Herein, a novel nanostructure based on cyclic aromatic polyimide with statistical star polymer structure was synthesized via the functionalization of the CuFe2O4 MNPs surface. The polymerization process on the functionalized surface of CuFe2O4 MNPs was performed with pyromellitic dianhydride and phenylenediamine derivatives. All analytical methods such as Fourier-transform infrared (FT-IR) spectroscopy, thermogravimetric (TG) analysis, X-ray diffraction (XRD) pattern, energy-dispersive X-ray (EDX), field-emission scanning electron microscope (FE-SEM), vibrating-sample magnetometer (VSM) were performed to characterize the structure of CuFe2O4@SiO2-polymer nanomagnetic. The cytotoxicity of CuFe2O4@SiO2-Polymer was investigated for biomedical application by MTT test. The results proved that this nanocmposite was biocompatible with HEK293T healthy cells. Also, the evaluation antibacterial property of CuFe2O4@SiO2-Polymer showed that its MIC in Gram-negative and Gram-positive bacteria were 500-1000 µg/mL, so it had antibacterial activity.

Bacterial production and biophysical characterization of a hard-to-fold scFv against myeloid leukemia cell surface marker, IL-1RAP.

BACKGROUND: Interleukin-1 receptor accessory protein (IL-1RAP) is one of the most promising therapeutic targets proposed for myeloid leukemia. Antibodies (Abs) specific to IL-1RAP could be valuable tools for targeted therapy of this lethal malignancy. This study is about the preparation of a difficult-to-produce single-chain variable fragment (scFv) construct against the membrane-bound isoform of human IL-1RAP using Escherichia coli (E. coli). METHODS: Different approaches were examined for refolding and characterization of the scFv. Binding activities of antibody fragments were comparatively evaluated using cell-based enzyme-linked immunosorbent assay (ELISA). Homogeneity and secondary structure of selected scFv preparation were analyzed using analytical size exclusion chromatography (SEC) and circular dichroism (CD) spectroscopy, respectively. The activity of the selected preparation was evaluated after long-term storage, repeated freeze-thaw cycles, or following incubation with normal and leukemic serum. RESULTS: Strategies for soluble expression of the scFv failed. Even with the help of Trx, ≥ 98% of proteins were expressed as inclusion bodies (IBs). Among three different refolding methods, the highest recovery rate was obtained from the dilution method (11.2%). Trx-tag substantially enhanced the expression level (18%, considering the molecular weight (MW) differences), recovery rate (˃1.6-fold), and binding activity (˃2.6-fold increase in absorbance450nm). The produced scFv exhibited expected secondary structure as well as acceptable bio-functionality, homogeneity, and stability. CONCLUSION: We were able to produce  21 mg/L culture functional and stable anti-IL-1RAP scFv via recovering IBs by pulse dilution procedure. The produced scFv as a useful targeting agent could be used in scheming new therapeutics or diagnostics for myeloid malignancies.

Magnetic chitosan-silk fibroin hydrogel/graphene oxide nanobiocomposite for biological and hyperthermia applications.

This work represents a biocompatible magnetic nanobiocomposite prepared by the composition of chitosan (CS) hydrogel, silk fibroin (SF), graphene oxide (GO), and Fe3O4 NPs. Terephthaloyl thiourea was applied as a cross-linking agent to cross-link the CS strings. The CS hydrogel/SF/GO/Fe3O4 nanobiocomposite with many characteristics, such as high structural uniformity, thermal stability, biocompatibility, and stability in an aqueous solution. Various characteristics of this novel magnetic nanobiocomposite were distinguished by FT-IR, EDX, FE-SEM, XRD, TGA, and VSM analysis. The FE-SEM images were taken to evaluate the size distribution of the magnetic nanoparticles (MNPs) between 39.9 and 73.3 nm as well. The performance of the prepared nanobiocomposite was assessed by the magnetic fluid hyperthermia process. Under the alternating magnetic field (AMF), the mean value of the specific absorption rate (SAR) was determined at 43.15 w/g.

Quality by Design in Downstream Process Development of Romiplostim

Background: Background: Downstream processing of therapeutic recombinant proteins expressed as the inclusion bodies (IBs) in E. coli is quite challenging. This study aimed to use the quality by design approach for developing the multi-step downstream process of a structurally complex therapeutic Fc-Peptide fusion protein, romiplostim. Methods: Methods: For development of a successful downstream process, risk analysis and experimental designs were used to characterize the most critical quality attributes (CQAs) and effects of process parameters on these quality attributes. Results: Results: The solubilization of IBs was optimized by design of experiment on three parameters with a focus on solubility yield, which resulted in >75% increase of the target protein solubilization. The pH of sample was identified as CQA in anion exchange chromatography that might have an impact on achieving >85% host cell proteins removal and >90% host cell DNA reduction. In the refolding step, process parameters were screened. Cystine/cysteine ratio, pH, and incubation time identified as CPPs were further optimized using Box-Behnken analysis, which >85% of the target protein was refolded. The design space for further purification step by HIC was mapped with a focus on high molecular weight impurities. After polishing by gel filtration, the final product's biological activity showed no statistically significant differences among the groups received romiplostim and Nplate®, as the reference product. Conclusions: Conclusion: This research presents a precise and exhaustive model for mapping the design space in order to describe and anticipate the link between the yield and quality of romiplostim and its downstream process parameters.

Fabrication of a magnetic alginate-silk fibroin hydrogel, containing halloysite nanotubes as a novel nanocomposite for biological and hyperthermia applications.

In this study, the main focus was on designing and synthesizing a novel magnetic nanobiocomposite and its application in hyperthermia cancer treatment. Regarding this aim, sodium alginate (SA) hydrogel with CaCl2 cross-linker formed and modified by silk fibroin (SF) natural polymer and halloysite nanotubes (HNTs), followed by in situ Fe3O4 magnetic nanoparticles preparation. No important differences were detected in red blood cells (RBCs) hemolysis, confirming the high blood compatibility of the treated erythrocytes with this nanobiocomposite. Moreover, the synthesized SA hydrogel/SF/HNTs/Fe3O4 nanobiocomposite does not demonstrate toxicity toward HEK293T normal cell line after 48 and 72 h. The anticancer property of SA hydrogel/SF/HNTs/Fe3O4 nanobiocomposites against breast cancer cell lines was corroborated. The magnetic saturation of the mentioned magnetic nanobiocomposite was 15.96 emu g-1. The specific absorption rate (SAR) was measured to be 22.3 W g-1 by applying an alternating magnetic field (AMF). This novel nanobiocomposite could perform efficiently in the magnetic fluid hyperthermia process, according to the obtained results.

Biological investigation of a novel nanocomposite based on functionalized graphene oxide nanosheets with pectin, silk fibroin and zinc chromite nanoparticles.

For biotechnology applications, a novel nanobiocomposite was synthesized based on modification of graphene oxide (GO) by extracted silk fibroin (SF), natural polymer pectin (Pec) and zinc chromite (ZnCr2O4) nanoparticles (NPs). The structure and properties of hybrid nanobiocomposite GO-Pec/SF/ZnCr2O4 such as thermal stability, less toxicity, biocompatibility, antibacterial, and biodegradable were proved by using field emission scanning electron microscope (FE-SEM), Fourier-transformed infrared (FT-IR), Energy dispersive X-ray spectroscopy (EDS), thermal gravimetric analysis (TGA), and X-Ray diffraction (XRD). According to the biological features of substances, the GO-Pec/SF/ZnCr2O4 nanobiocomposite shows perfect results in MTT (83.71 %) and Hemolysis (16.52 %) assays. accordingly, mentioned properties of this nanobiocomposite can be used as a scaffold for medical applications.

Functionalized graphene oxide nanosheets with folic acid and silk fibroin as a novel nanobiocomposite for biomedical applications.

In this paper, a novel graphene oxide-folic acid/silk fibroin (GO-FA/SF) nanobiocomposite scaffold was designed and fabricated using affordable and non-toxic materials. The GO was synthesized using the hummer method, covalently functionalized with FA, and then easily conjugated with extracted SF via the freeze-drying process. For characterization of the scaffold, several techniques were employed: Fourier-transform infrared (FT-IR), field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray (EDX), and thermogravimetric analysis (TGA). The cell viability method, hemolysis, and anti-biofilm assays were performed, exploring the biological capability of the nanobiocomposite. The cell viability percentages were 96.67, 96.35 and 97.23% for 24, 48, and 72 h, respectively, and its hemolytic effect was less than 10%. In addition, it was shown that this nanobiocomposite prevents the formation of Pseudomonas aeruginosa biofilm and has antibacterial activity.

COVID-19: A systematic review and update on prevention, diagnosis, and treatment.

Since the rapid onset of the COVID-19 or SARS-CoV-2 pandemic in the world in 2019, extensive studies have been conducted to unveil the behavior and emission pattern of the virus in order to determine the best ways to diagnosis of virus and thereof formulate effective drugs or vaccines to combat the disease. The emergence of novel diagnostic and therapeutic techniques considering the multiplicity of reports from one side and contradictions in assessments from the other side necessitates instantaneous updates on the progress of clinical investigations. There is also growing public anxiety from time to time mutation of COVID-19, as reflected in considerable mortality and transmission, respectively, from delta and Omicron variants. We comprehensively review and summarize different aspects of prevention, diagnosis, and treatment of COVID-19. First, biological characteristics of COVID-19 were explained from diagnosis standpoint. Thereafter, the preclinical animal models of COVID-19 were discussed to frame the symptoms and clinical effects of COVID-19 from patient to patient with treatment strategies and in-silico/computational biology. Finally, the opportunities and challenges of nanoscience/nanotechnology in identification, diagnosis, and treatment of COVID-19 were discussed. This review covers almost all SARS-CoV-2-related topics extensively to deepen the understanding of the latest achievements (last updated on January 11, 2022).

Computational Engineering of Protein L to Achieve an Optimal Affinity Chromatography Resin for Purification of Antibody Fragments.

Protein L affinity chromatography is a useful method for the purification of antibody fragments containing kappa light chains. In affinity chromatography, increasing the binding affinity leads to increased product purity, recovery, and dynamic binding capacity (DBC). In this study, molecular docking and molecular dynamics simulation techniques were used to design the engineered Protein L with higher affinity to the kappa light chain. Each engineered ligand was produced as a recombinant protein and coupled to a solid matrix. The purity, recovery, and DBC of the engineered resins were evaluated and then compared to those of a commercially available resin. The results showed important parameters for engineering more efficient Protein L ligands for affinity chromatography.

Novel magnetic organic-inorganic hybrids based on aromatic polyamides and ZnFe2O4 nanoparticles with biological activity.

Magnetic nanoparticles were creatively selected as stable, inexpensive, biodegradable, facile recoverable, and functionalizable supports for a variety of synthetic and natural polymers. Herein, for the first time, aromatic polyamide was synthesized on the magnetic core of zinc iron oxide (ZnFe2O4). Terephthaloyl chloride and derivations of phenylenediamine were employed as monomers in this polymerization process. The toxicity of the synthesized hybrid at the highest concentration (1000 µg/ml) is 13.65% and on the other hand, the cell viability percentage is 86.35%. So, the prepared hybrid is biocompatible and non-toxic to Hu02 cells. Also, it has antibacterial ability against gram-positive and gram-negative bacteria. Because the results show that the minimum inhibitory concentration (MIC) of the synthesized polymer for bacteria such as Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa ATCC 27853 is in the range of 500-1000 µg/ml. Moreover, the hemolytic effect of ZnFe2O4 based hybrid was below 9% at the concentration of 1000 µg/ml. Therefore, it is compatible with red blood cells.