Showing papers on "Bovine serum albumin published in 2021"

Comparison of non-covalent binding interactions between three whey proteins and chlorogenic acid: Spectroscopic analysis and molecular docking

Abstract: Non-covalent binding interactions between three whey proteins β-lactoglobulin (β-Lg), α-lactalbumin (α-La) and bovine serum albumin (BSA) with chlorogenic acid (CA) were investigated using spectroscopic analysis and molecular docking Fluorescence study showed that CA quenched the fluorescence of three whey proteins through static mode The binding number was equal to 1 for three proteins and binding affinity in declined order was: α-La > β-Lg > BSA Thermodynamic parameters revealed contribution of hydrophobic force in three systems Fluorescence resonance energy transfer (FRET) measurements indicated that energy transfer occurs between three proteins and CA in probability of α-La > BSA > β-Lg Variation in surface charge indicated the involvement of electrostatic interaction Surface hydrophobicity (H0) were declined with decrease degree of α-La > β-Lg > BSA α-La and β-Lg were unfolded with more flexible structure while BSA skeleton was more compact after interacting with CA Modeling study revealed that the most likely binding sites for the three proteins were outer surface, cleft and subdomain I for β-Lg, α-La and BSA respectively Docking results also suggested the contribution of hydrophobic interaction and hydrogen bond (β-Lg, α-La) for formation of molecular nano complexes between whey proteins and CA

The Most Active Oxidase-Mimicking Mn 2 O 3 Nanozyme for Biosensor Signal Generation

Abstract: Oxidase-mimicking nanozymes are more desirable than peroxidase-mimicking ones since H2 O2 can be omitted. However, only a few nanomaterials are known for oxidase-like activities. In this work, we compared the activity of Mn2 O3 , Mn3 O4 and MnO2 and found that Mn2 O3 had the highest oxidase activity. Interestingly, the activity of Mn2 O3 was even inhibited by H2 O2 . The oxidase-like activity of Mn2 O3 was not much affected by the presence of proteins such as bovine serum albumin (BSA), but the physisorption of antibodies to Mn2 O3 was not strong enough to withstand the displacement by BSA. We then treated Mn2 O3 with 3-aminopropyltriethoxysilane to graft an amine group, which was used to conjugate antibodies using glutaraldehyde as a crosslinker. A one-step indirect competitive ELISA (icELISA) was developed for the detection of isocarbophos, and an IC50 of 261.7 ng/mL was obtained, comparable with the results of the standard two-step assay using horseradish peroxidase (HRP)-labeled antibodies. This assay has the advantage of significant timesaving for rapid detection of large amounts of samples. This work has discovered a highly efficient oxidase-mimicking nanozyme useful for various nano- and analytical applications.

Bovine Serum Albumin-Cross-Linked Polyaniline Nanowires for Ultralow Fouling and Highly Sensitive Electrochemical Protein Quantification in Human Serum Samples.

Abstract: Biofouling represents a serious challenge for the assaying of disease markers with various biosensors in complex biological samples due to the accompanied nonspecific protein adsorption. Herein, a highly sensitive and antifouling biosensing interface was constructed based on a cost-effective inert protein bovine serum albumin (BSA) cross-linked with polyaniline nanowires (PANI-NWs). Compared with the physically adsorbed BSA that was commonly used to block nonspecific adsorption or binding of proteins, the cross-linked BSA exhibited a significantly enhanced antifouling capability. The BSA/PANI-NW-modified electrode interface possessed excellent antifouling capability and electrochemical activity owing to the presence of the cross-linked BSA and the conducting polymer polyaniline. With further immobilization of the peptide aptamer for immunoglobulin G (IgG) recognition onto the BSA/PANI-NW interface, an electrochemical biosensor with excellent selectivity and sensitivity was prepared. The IgG biosensor possessed a linear range from 1.0 ng mL-1 to 10 μg mL-1 and a low detection limit of 0.27 ng mL-1, and it was capable of assaying IgG in complex human serum samples with acceptable accuracy when compared with the assay results obtained using commercial enzyme-linked immunosorbent assay kits. It is expected that the unique BSA-cross-linked conducting polymers can be used for the construction of various electrochemical sensors and biosensors that can be applied in complex biological media.

Probing the Interaction of Bovine Serum Albumin with Copper Nanoclusters: Realization of Binding Pathway Different from Protein Corona

Abstract: With an aim to understand the interaction mechanism of bovine serum albumin (BSA) with copper nanoclusters (CuNCs), three different types CuNCs having chemically different surface ligands, namely, tannic acid (TA), chitosan, and cysteine (Cys), have been fabricated, and investigations are carried out in the absence and presence of protein (BSA) at ensemble-averaged and single-molecule levels. The CuNCs, capped with different surface ligands, are consciously chosen so that the role of surface ligands in the overall protein-NCs interactions is clearly understood, but, more importantly, to find whether these CuNCs can interact with protein in a new pathway without forming the "protein corona", which otherwise has been observed in relatively larger nanoparticles when they are exposed to biological fluids. Analysis of the data obtained from fluorescence, ζ-potential, and ITC measurements has clearly indicated that the BSA protein in the presence of CuNCs does not attain the binding stoichiometry (BSA/CuNCs > 1) that is required for the formation of "protein corona". This conclusion is further substantiated by the outcome of the fluorescence correlation spectroscopy (FCS) study. Further analysis of data and thermodynamic calculations have revealed that the surface ligands of the CuNCs play an important role in the protein-NCs binding events, and they can alter the mode and thermodynamics of the process. Specifically, the data have demonstrated that the binding of BSA with TA-CuNCs and Chitosan-CuNCs follows two types of binding modes; however, the same with Cys-CuNCs goes through only one type of binding mode. Circular dichroism (CD) measurements have indicated that the basic structure of BSA remains almost unaltered in the presence of CuNCs. The outcome of the present study is expected to encourage and enable better application of NCs in biological applications.

Highly Reproducible and Sensitive Electrochemiluminescence Biosensors for HPV Detection Based on Bovine Serum Albumin Carrier Platforms and Hyperbranched Rolling Circle Amplification.

Abstract: Most DNA-based electrochemiluminescence (ECL) biosensors are established through the self-assembly of thiolated single-stranded DNA (ssDNA) probes on the Au electrode surface. Because of this random assembly process, a significant discrepancy exists in the distribution of a modified DNA film on different electrodes, which greatly affects the reproducibility of a biosensor. In this study, a porous bovine serum albumin (BSA) layer was first modified on the electrode surface, which can improve the position distribution and spatial orientation of the self-assembly ssDNA probe. It was then coupled with hyperbranched rolling circle amplification to develop the high-reproducibility-and-sensitivity ECL biosensor for human papillomavirus 16 E6 and E7 oncogene detection. In the presence of the target DNA, the surface of the electrode accumulates abundant amplified products through reaction, which contain double-stranded DNA (dsDNA) fragments of different lengths, followed by plentiful dichlorotris (1,10-phenanthroline) ruthenium(II) hydrate (Ru(phen)32+, acting as an ECL indicator) insertion into grooves of dsDNA fragments, and a strong signal can be detected. There is a linear relationship between the signal and the target concentration range from 10 fM to 15 pM, and the detection limit is 7.6 fM (S/N = 3). After the BSA modification step, the relative standard deviation was reduced from 9.20 to 3.96%, thereby achieving good reproducibility. The proposed ECL strategy provides a new method for constructing high-reproducibility-and-sensitivity ECL biosensors.

Interactions of blueberry anthocyanins with whey protein isolate and bovine serum protein: Color stability, antioxidant activity, in vitro simulation, and protein functionality

Abstract: The interaction of proteins with anthocyanins (ACNs) gives rise to significant improvements in their functional properties. To explore the interactions between whey protein isolate (WPI)/bovine serum albumin (BSA) and blueberry ACNs, light (5000 Lux, 7 days), heat (85 °C, 2 h), sucrose (0.25 mg/mL), and vitamin C (0.15 mg/mL) were used to accelerate the degradation of ACNs. Through the observation of color changes and variations in the free radical scavenging rates, WPI and BSA were found to protect the ACNs to varying degrees. Moreover, WPI/BSA exhibited increased solubilities, foaming and emulsifying properties, and changed thermal stabilities and protein structures, in addition to decreased surface hydrophobicities and zeta potentials. Furthermore, the addition of WPI/BSA increased the number of ACN monomers in a simulated digestion system, as determined by high-performance liquid chromatography-mass spectrometry. Therefore, by studying the interactions between proteins and ACNs, we can provide a strategy and theoretical basis for determining the steady-state of ACNs in blueberry products.

Effect of proteins on the oxidase-like activity of CeO2 nanozymes for immunoassays.

Abstract: Having the benefits of low cost, excellent stability and tolerance to extreme conditions, nanozymes are a potential alternative of horseradish peroxidase (HRP) or other enzymes for bioanalytical chemistry, especially immunoassays. CeO2 nanoparticles have oxidase-mimicking activity and can avoid the use of unstable H2O2. For robust assays, the effect of proteins on the activity of CeO2 needs to be carefully studied. Herein, we studied the adsorption and desorption of bovine serum albumin (BSA) from CeO2. The CeO2 nanoparticles exhibited a higher protein adsorption capacity compared to the other tested metal oxide nanoparticles. Although the oxidase-like activity of CeO2 was inhibited by BSA, low concentrations of phosphate and fluoride ions boosted the activity of protein-capped CeO2. CeO2 was still active under strong acidic conditions and at high temperature, while HRP lost its activity. For immunoassay development, we covered CeO2 with an amine-modified silane for covalent conjugation to antibodies. A one-step indirect competitive ELISA for fenitrothion was developed, and an IC50 value of 35.6 ng mL-1 and a limit of detection of 2.1 ng mL-1 were obtained.

Investigating the mechanism of binding of nalidixic acid with deoxyribonucleic acid and serum albumin: a biophysical and molecular docking approaches

Abstract: Nalidixic acid is a bacterial DNA gyrase inhibitor and the first member of the synthetic quinolone antibiotics. It is used in the treatment of various infectious diseases like urinary tract infecti...

Bulk Phase Behavior vs Interface Adsorption: Specific Multivalent Cation and Anion Effects on BSA Interactions.

Abstract: Proteins are ubiquitous and play a critical role in many areas from living organisms to protein microchips. In humans, serum albumin has a prominent role in the foreign body response since it is the first protein which will interact with, e.g., an implant or stent. In this study, we focused on the influence of salts (i.e., different cations (Y3+, La3+) and anions (Cl-, I-) on bovine serum albumin (BSA) in terms of its bulk behavior as well as the role of charges for protein adsorption at the solid-liquid interface in order to understand and control the underlying molecular mechanisms and interactions. This is part of our group's effort to gain a deeper understanding of protein-protein and protein-surface interactions in the presence of multivalent ions. In the bulk, we established two new phase diagrams and found not only multivalent cation-triggered phase transitions, but also a dependence of the protein behavior on the type of anion. The attractive interactions between proteins were observed to increase from Cl- < NO3- < I-, resulting in iodide preventing re-entrant condensation and promoting liquid-liquid phase separation in bulk. Using ellipsometry and a quartz-crystal microbalance with dissipation (QCM-D), we obtained insight into the growth of the protein adsorption layer. Importantly, we found that phase transitions at the substrate can be triggered by certain interface properties, whether they exist in the bulk solution or not. Through the use of a hydrophilic, negatively charged surface (native silica), the direct binding of anions to the interface was prevented. Interestingly, this led to re-entrant adsorption even in the absence of re-entrant condensation in bulk. However, the overall amount of adsorbed protein was enhanced through stronger attractive protein-protein interactions in the presence of iodide salts. These findings illustrate how carefully chosen surface properties and salts can directly steer the binding of anions and cations, which guide protein behavior, thus paving the way for specific/triggered protein-protein, protein-salt, and protein-surface interactions.