Radioprotective Role of Vitamins C and E against the Gamma Ray-Induced Damage to the Chemical Structure of Bovine Serum Albumin.

Radioprotective effects of vitamin C and vitamin E as a water-soluble and a lipid-soluble agent, respectively, were investigated at the molecular level during the imposition of gamma radiation-induced structural changes to bovine serum albumin (BSA) at the therapeutic dose of 3 Gy. Secondary and tertiary structural changes of control and irradiated BSA samples were investigated using circular dichroism and fluorescence spectroscopy. The preirradiation tests showed nonspecific and reversible binding of vitamins C and E to BSA. Secondary and tertiary structures of irradiated BSA considerably changed in the absence of the vitamins. Upon irradiation, α-helices of BSA transitioned to beta motifs and random coils, and the fluorescence emission intensity decreased relative to nonirradiated BSA. In the presence of the vitamins C or E, however, the irradiated BSA was protected from these structural changes caused by reactive oxygen species (ROS). The two vitamins exhibited different patterns of attachment to the protein surface, as inspected by blind docking, and their mechanisms of protection were different. The hydrophilicity of vitamin C resulted in the predominant scavenging of ROS in the solvent, whereas hydrophobic vitamin E localized on the nonpolar patches of the BSA surface, where it did not only form a barrier for diffusing ROS but also encountered them as an antioxidant and neutralized them thanks to the moderate BSA binding constant. Very low concentrations of vitamins C or E (0.005 mg/mL) appear to be sufficient to prevent the oxidative damage of BSA.

A high-sensitivity electrochemical immunosensor based on mobile crystalline material-41-polyvinyl alcohol nanocomposite and colloidal gold nanoparticles.

A novel competitive immunosensor was developed as a model system using anti-human serum albumin (HSA)-conjugated gold nanoparticles (AuNPs) as an electrochemical label and mobile crystalline material-41 (MCM-41)-polyvinyl alcohol (PVA) mesoporous nanocomposite as an immobilization platform. However, no attempt has yet been made to use the MCM-41 as the supporting electrolyte for the electrosynthesis of nonconducting polymer nanocomposite. This hybrid membrane was evaluated extensively by using field emission scanning electron microscopy (FESEM), cyclic voltammetry (CV), and differential pulse voltammetry (DPV) to determine its physicochemical and electrochemical properties in immunosensor application. FESEM revealed an appropriate and stable attachment between HSA and MCM-41 and also a dense layer deposition of MCM-41-HSA-PVA film onto the electrode surfaces. DPV was developed for quantitative determination of antigen in biological samples. A decrease in DPV responses was observed with increasing concentrations of HSA in standard and real samples. In optimal conditions, this immunosensor based on MCM-41-PVA nanocomposite film could detect HSA in a high linear range (0.5-200 µg ml⁻¹) with a low detection limit of 1 ng ml⁻¹. The proposed method showed acceptable reproducibility, stability, and reliability and could also be applied to detect the other antigens.

Magnetic nanocomposite of anti-human IgG/COOH-multiwalled carbon nanotubes/Fe3O4 as a platform for electrochemical immunoassay.

An electrochemical immunosensing method was developed based on a magnetic nanocomposite. The multiwalled carbon nanotubes (MWCNTs) were treated with nitric acid to produce carboxyl groups at the open ends. Then, Fe3O4 nanoparticles were deposited on COOH-MWCNTs by chemical coprecipitation of Fe²âº and Fe³âº salts in an alkaline solution. Goat anti-human IgG (anti-hIgG) was covalently attached to magnetic nanocomposite through amide bond formation between the carboxylic groups of MWCNTs and the amine groups of anti-hIgG. The prepared bio-nanocomposite was used for electrochemical sensing of human tetanus IgG (hIgG) as a model antigen. The anti-hIgG magnetic nanocomposite was fixed on the surface of a gold plate electrode using a permanent magnet. The hIgG was detected using horseradish peroxidase (HRP)-conjugated anti-hIgG in a sandwich model. Electrochemical detection of hIgG was carried out in the presence of H2O2 and KI as substrates of HRP. Using this method, hIgG was detected in a concentration range from 30 to 1000 ng ml⁻¹ with a correlation coefficient of 0.998 and a detection limit of 25 ng ml⁻¹ (signal/noise=3). The designed immunosensor was stable for 1 month.

Development of urinary albumin immunosensor based on colloidal AuNP and PVA.

The development of immunosensors with high sensitivity and specificity in detecting the pathogenic or physiologically relevant molecules in the body, offers a powerful opportunity in early diagnosis and treatment of diseases. In this study, we developed a new competitive immunosensor with employing antibody (Ab) labeled AuNP (Ab-AuNP) and PVA modified screen-printed carbon electrode (SPCE) surface to detect the urine albumin. Field emission scanning electron microscopy (FE-SEM) of modified electrode showed a suitable and stable attachment between HSA antigen- mAb and AuNP. Cyclic voltammetric (CV) method demonstrated that modification process was well performed. Electrochemical measurements including differential pulse voltammetry (DPV) and square wave voltammetry (SWV) were employed for quantitative antigen detection. The electrochemical measurements performed with other proteins mixed with samples demonstrated a high specificity and selectivity for this biosensor in detecting the HSA. In optimal conditions, the immunosensor could detect HSA in a high linear range (from 2.5 to 200 µg/mL) with a low detection limit of 25 ng/mL. This new strategy could be improved and applied to detect the other antigen.