ELISA Sisteminde Protein-İnorganik Hibrit Konjugatın Kullanılabilirliği

yokIn the Enzyme-linked immunosorbent assay method (ELISA), it is important to increase the immobilization efficiency of the functional molecules on the microplate and increase the signal-to-noise ratio in order to detect the target molecules with high sensitivity and selectivity. For this purpose, antibody functionalized materials can generate remarkable signal amplification with high enzyme capacity by using conventional immobilization methods. Recently, hybrid structures containing protein/enzyme and Cu3(PO4)2, which is a different enzyme immobilization method, have higher catalytic activity compared to the free form of protein-containing molecules as they have hierarchical microstructures and form large active surface areas. In this study; using protein-inorganic hybrid structure synthesis method, hybrid functionalized conjugate systems with enzyme, antibody and Cu3(PO4)2 were synthesized all together and characterization of the resulting structures was performed by SEM, EDX, XRD and FTIR analysis. When the findings obtained from the researches were evaluated, the hybrid nano structure showed 183.5 EU/mg peroxidase activity and free HRP enzyme showed 59.01 EU/mg activity. The performance of the ELISA system was measured using hybrid conjugate constructs prepared using various TNF-alpha specific antibodies at a -1 concentration of 5-1000 g mL . The performance of hybrid conjugate structure containing multiple organic molecules in ELISA system was higher than other structures. This method is a highly practical method that can replace enzyme-labeled antibody method in ELISA.yo

Kudret Narı (Momordica charantia Descourt.) Meyvesinden Saflaştırılan Peroksidaz Enzimi Kullanılarak Hibrit Nano Çiçekler Sentezlenmesi ve Direct Blue 1 Gideriminde Kullanılabilirlikleri

.Peroxidase enzymes are purified from different plant sources are used efficiently for the removal of dyes in industrial wastes. The fruit of bitter gourd (Momordica charantia), an inexpensive and easily accessible vegetable, is an important source of peroxidase. In this study, total protein content was found to be 0.485 mg/mL and peroxidase activity was found to be 2360.9 EU/mg as a result of 50% protein precipitation made from green bitter gourd. However; total protein amount was 0.232 mg/mL and free peroxidase activity was determined as 7719.30 EU/mg as a result of 60% protein precipitation made from ripe bitter gourd. Peroxidase enzymes which were purified from bitter gourd in different growth stages under optimum conditions showed higher enzymatic activity compared to free forms when immobilized via enzyme-inorganic hybrid nanoflower synthesis method. The highest peroxidase activity was seen in mature fruit and hybrid nanoflower form (19661, 6 EU/mg). In addition, the usability of hybrid nanoflowers was investigated compared to the free purified peroxidase for removal of Direct Blue 1 dye widely used in textile industry. It was determined that hybrid nanoflower form synthesized especially by using ripe bitter gourd peroxidase had more dye removal.

A new approach for green synthesis and characterization of Artemisia L. (Asteraceae) genotype extracts-Cu2 nanocomplexes (nanoflower) and their effective antimicrobial activity

.In this study, we have demonstrated the fabrication of novel organic-inorganic nanobio-antimicrobial agents called “nanoflowers” (NFs) and elucidate the increase in the antimicrobial activity of NFs. This is the first report that the NFs were formed of plant extracts as the organic components and copper (II) ions (Cu2+) as the inorganic component. The Artemisia L. (Asteraceae) methanol extracts from three genotypes including A. absinthium L. (Aa), A. vulgaris L. (Av) and A. ludoviciana Nutt. (Al) were selected in the NF synthesis. The effect of the plant extract concentrations on the morphology of NFs was examined. Most regular and uniform flower-shaped morpholo- gies were observed when a concentration of 0.1 mg mL-1 plant extract was used in the synthesis of NFs. The syntesized NFs were characterized with several techniques such as scanning electron microscopy (SEM), Fourier transform infrared spectrometer (FT-IR), energy-dispersive X-ray (EDX) and X-ray diffraction analysis (XRD). The NFs exhibited much antimicrobial activity against the pathogens even at low concentrations compared to the extracts. The MICs and MBCs values for NFs were found to be range between 0.4 to 40 μg mL-1 and 40 to 400 μg mL-1 while those values for Aa, Av and Al extracts were ranged from 500-2000 μg mL-1 and 1000-4000 μg mL-1 for the studied pathogens, respectively.

Evaluating the activity and stability of sonochemically produced hemoglobin-copper hybrid nanoflowers against some metallic ions, organic solvents, and inhibitors

The disadvantage of the conventional protein-inorganic hybrid nanoflower production method is the long incubation period of the synthesis method. This period is not suitable for practical industrial use. Herein, protein-inorganic hybrid nanoflowers were synthesized using hemoglobin and copper ion by fast sonication method for 10 min. The synthesized nanoflowers were characterized via scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction, and Fouirer-transform infrared spectroscopy. The activity and stability of the nanoflowers in the presence of different metal ions, organic solvents, inhibitors, and storage conditions were also evaluated by comparing with free hemoglobin. According to obtained results, the optimum pH and temperatures of both hybrid nanoflower and free hemoglobin were pH 5 and 40 °C, respectively. At all pH levels, nanoflower was more stable than free protein and it was also more stable than the free hemoglobin at temperatures ranging between 50 °C and 80 °C. The free protein lost more than half of its activity in the presence of acetone, benzene, and N,N-dimethylformamide, while the hybrid nanoflower retained more than 70% of its activity for 2 h at 40 °C. The hybrid nanoflower activity was essentially increased in the presence of Ca2+, Zn2+, Fe2+, Cu2+ and Ni2+ (132%, 161%, 175%, 185% and 106%, respectively) at 5 mM concentration. The nanoflower retained more than 85% of its initial activity in the presence of all inhibitors. In addition, it retained all its activity for 3 days under different storage conditions, unlike free hemoglobin. The results demonstrated that new hybrid nanoflowers may be promising in different biotechnological applications such as catalytic biosensors and environmental or industrial catalytic processes.Scientific Research Projects Coordination Unit for technical infrastructure of Nevsehir Haci Bektas Veli University Scientific Research Projects Coordination Unit for technical infrastructure of Kafkas University Scientific Research Projects Coordination Unit for technical infrastructure of Igdir Universit

Hemoglobin-metal2+ phosphate nanoflowers with enhanced peroxidase-like activities and their performance in visual detection of hydrogen peroxide

.Hemoglobin (Hgb)–metal 2+ 2+ 2+ phosphate nanoflowers (Hgb–X -Nfs) were synthesized using Co 2+ , Zn , 2+ 2+ Ca , and Fe separately as inorganic components, to generate a visual hydrogen peroxide (H2O2) biosensor for the first time. While there was no flower-like structure using Ca2+ and Fe2+ ions, the average diameters of nanoflowers obtained using Co2+ and Zn2+ were about 7–8 mm and 1.2 mm, respectively. Their encapsulation efficiencies were 85% and 90%, respectively. The peroxidase-like activities of Hgb–X2+-Nfs were evaluated against the ABTS substrate, compared to free hemoglobin. The obtained Hgb–X2+-Nfs were used as a biocatalyst to generate visual and spectrometric biosensors. The working range that can be detected visually was 0.005–0.667 mM in both hybrid nanoflowers, while the working range that can be detected spectrometrically was 0.00017–0.667 mM for Hgb–Co2+-Nfs and 0.0017–0.667 mM for Hgb–Zn2+-Nfs. The proposed biosensing methods were successfully applied for the determination of hydrogen peroxide in tap water and milk samples, with recoveries ranging from 95.23% to 107.01%. The findings of this study can be used for the production of new hybrid nanoflowers incorporating hemoglobin and various metal ions for use as potential biosensors. In addition, the proposed biosensors have the potential to provide a successful analytical tool in many areas of analysis, such as pharmaceutical, clinical, food and environmental.yo

ELISA SİSTEMİNDE PROTEİN-İNORGANİK HİBRİT KONJUGATIN KULLANILABİLİRLİĞİ

yokIn the Enzyme-linked immunosorbent assay method (ELISA), it is important to increase the immobilization efficiency of the functional molecules on the microplate and increase the signal-to-noise ratio in order to detect the target molecules with high sensitivity and selectivity. For this purpose, antibody functionalized materials can generate remarkable signal amplification with high enzyme capacity by using conventional immobilization methods. Recently, hybrid structures containing protein/enzyme and Cu3(PO4)2, which is a different enzyme immobilization method, have higher catalytic activity compared to the free form of protein-containing molecules as they have hierarchical microstructures and form large active surface areas. In this study; using protein-inorganic hybrid structure synthesis method, hybrid functionalized conjugate systems with enzyme, antibody and Cu3(PO4)2 were synthesized all together and characterization of the resulting structures was performed by SEM, EDX, XRD and FTIR analysis. When the findings obtained from the researches were evaluated, the hybrid nano structure showed 183.5 EU/mg peroxidase activity and free HRP enzyme showed 59.01 EU/mg activity. The performance of the ELISA system was measured using hybrid conjugate constructs prepared using various TNF-alpha specific antibodies at a -1 concentration of 5-1000 g mL . The performance of hybrid conjugate structure containing multiple organic molecules in ELISA system was higher than other structures. This method is a highly practical method that can replace enzyme-labeled antibody method in ELISA.yo