Synergistic integration of Ni-metal organic framework/SnS2nanocomposite and nickel foam electrode for ultrasensitive and selective electrochemical detection of albumin in simulated human blood serum.

Albumin is a vital blood protein responsible for transporting metabolites and drugs throughout the body and serves as a potential biomarker for various medical conditions, including inflammatory, cardiovascular, and renal issues. This report details the fabrication of Ni-metal organic framework/SnS2nanocomposite modified nickel foam electrochemical sensor for highly sensitive and selective non enzymatic detection of albumin in simulated human blood serum samples. Ni-metal organic framework/SnS2nanocomposite was synthesized using solvothermal technique by combining Ni-metal-organic framework (MOF) with conductive SnS2leading to the formation of a highly porous material with reduced toxicity and excellent electrical conductivity. Detailed surface morphology and chemical bonding of the Ni-MOF/SnS2nanocomposite was studied using scanning electron microscopy, transmission electron microscopy, Fourier transform infra-red, and Raman analysis. The Ni-MOF/SnS2nanocomposite coated on Ni foam electrode demonstrated outstanding electrochemical performance, with a low limit of detection (0.44µM) and high sensitivity (1.3µA/pM/cm2) throughout a broad linear range (100 pM-10 mM). The remarkable sensor performance is achieved through the synthesis of a Ni-MOF/SnS2nanocomposite, enhancing electrocatalytic activity for efficient albumin redox reactions. The enhanced performance can be attributed due to the structural porosity of nickel foam and Ni-metal organic framework, which favours increased surface area for albumin interaction. The presence of SnS2shows stability in acidic and neutral solutions due to high surface to volume ratio which in turn improves sensitivity of the sensing material. The sensor exhibited commendable selectivity, maintaining its performance even when exposed to potential interfering substances like glucose, ascorbic acid, K+, Na+, uric acid, and urea. The sensor effectively demonstrates its accuracy in detecting albumin in real samples, showcasing substantial recovery percentages of 105.1%, 110.28%, and 91.16%.

Thiourea Functionalised Receptor for Selective Detection of Mercury Ions and its Application in Serum Sample.

A thiourea functionalised fluorescent probe 1-phenyl-3-(pyridin-4-yl)thiourea was synthesized and utilised as a fluorescent turn-on chemosensor for the selective recognition of Hg2+ ion over competitive metal ions including Na+, Mn2+, Li+, Cr2+, Ni2+, Ca2+, Cd2+, Mg2+, K+, Co2+, Cu2+, Zn2+, Al3+ and Fe2+ ions based on the inter-molecular charge transfer (ICT). Intriguingly, the receptor demonstrated unique sensing capabilities for Hg2+ in DMSO: H2O (10:90, v/v). The addition of Hg2+ ions to the sensor resulted in a blue shift in the absorption intensity and also enhancement in fluorescence intensity at 435 nm. Fluorescence emission intensity increased linearly with Hg2+ concentration ranging from 0 to 80 µL. The detection limit and binding constant were determined as 0.134 × 10-6 M and 1.733 × 107 M-1, respectively. The sensing behavior of Hg2+ was further examined using DLS, SEM and FTIR. The probe could detect Hg2+ ions across a wide pH range. Furthermore, the receptor L demonstrated good sensing performance for Hg2+ in bovine serum albumin and actual water samples.

3D Nanolithography by Means of Lipid Ink Spreading Inhibition.

While patterning 2D metallic nanostructures are well established through different techniques, 3D printing still constitutes a major bottleneck on the way to device miniaturization. In this work a fluid phase phospholipid ink is used as a building block for structuring with dip-pen nanolithography. Following a bioinspired approach that relies on ink-spreading inhibition, two processes are presented to build 2D and 3D metallic structures. Serum albumin, a widely used protein with an innate capability to bind to lipids, is the key in both processes. Covering the sample surface with it prior to lipid writing, anchors lipids on the substrate, which ultimately allows the creation of highly stable 3D lipid-based scaffolds to build metallic structures.

BSA modification of bacterial surface: a promising anti-cancer therapeutic strategy.

BACKGROUND: Attenuated live bacterial therapy and medical BSA materials have their own advantages in anti-cancer research, and their combination is expected to overcome some of the disadvantages of conventional anti-cancer therapeutics. METHODS AND OBJECTIVE: Utilizing the high affinity between biotin and streptavidin, BSA modification on the surface of Escherichia coli (E. coli) was achieved. Then, the adhesion and targeting abilities of BSA modified E. coli was explored on different bladder cancer cells, and the underlying mechanism was also investigated. RESULTS: BSA modification on the surface of E. coli enhances its ability to adhere and target cancer cells, and we speculate that these characteristics are related to the expression of SPARC in different bladder cancer cell lines. CONCLUSION: BSA and live bacteria have their own advantages in anti-cancer research. In this study, we found that E. coli surface-modified by BSA had stronger adhesion and targeting effects on bladder cancer cells with high expression of SPARC. These findings pave the way for the future studies exploring the combination of BSA combined with live bacteria for cancer therapy.

pH-Dependent Changes in Structural Stabilities of Bt Cry1Ac Toxin and Contrasting Model Proteins following Adsorption on Montmorillonite.

The environmental fate of insecticidal Cry proteins, including time-dependent conservation of biological properties, results from their structural stability in soils. The complex cascade of reactions involved in biological action requires Cry proteins to be in solution. However, the pH-dependent changes in conformational stability and the adsorption-desorption mechanisms of Cry protein on soil minerals remain unclear. We used Derjaguin-Landau-Verwey-Overbeek (DLVO) calculation and differential scanning calorimetry to interpret the driving forces and structural stabilities of Cry1Ac and two contrasting model proteins adsorbed by montmorillonite. The structural stability of Cry1Ac is closer to that of the "hard" protein, α-chymotrypsin, than that of the "soft" bovine serum albumin (BSA). The pH-dependent adsorption of Cry1Ac and α-chymotrypsin could be explained by DLVO theory, whereas the BSA adsorption deviated from it. Patch-controlled electrostatic attraction, hydrophobic effects, and entropy changes following protein unfolding on a mineral surface could contribute to Cry1Ac adsorption. Cry1Ac, like chymotrypsin, was partly denatured on montmorillonite, and its structural stability decreased with an increase in pH. Moreover, small changes in the conformational heterogeneity of both Cry1Ac and chymotrypsin were observed following adsorption. Conversely, adsorbed BSA was completely denatured regardless of the solution pH. The moderate conformational rearrangement of adsorbed Cry1Ac may partially explain why the insecticidal activity of Bt toxin appears to be conserved in soils, albeit for a relatively short time period.

Macromolecular Crowding and DNA: Bridging the Gap between In Vitro and In Vivo.

The cellular environment is highly crowded, with up to 40% of the volume fraction of the cell occupied by various macromolecules. Most laboratory experiments take place in dilute buffer solutions; by adding various synthetic or organic macromolecules, researchers have begun to bridge the gap between in vitro and in vivo measurements. This is a review of the reported effects of macromolecular crowding on the compaction and extension of DNA, the effect of macromolecular crowding on DNA kinetics, and protein-DNA interactions. Theoretical models related to macromolecular crowding and DNA are briefly reviewed. Gaps in the literature, including the use of biologically relevant crowders, simultaneous use of multi-sized crowders, empirical connections between macromolecular crowding and liquid-liquid phase separation of nucleic materials are discussed.

Research and Application of Kupffer Cell Thresholds for BSA Nanoparticles.

Over the past decade, the dose of nanoparticles given to solid tumors has remained at a median of 0.7% of the injected dose. Most nanoparticles are trapped in a mononuclear phagocyte system (MPS), of which 85% are Kupffer cells. In our study, threshold doses of bovine serum albumin (BSA) nanoparticles were investigated for the uptake of Kupffer cells in vitro and in vivo. The antitumor effect and safety of albumin-bound paclitaxel (ABP) were improved by using threshold doses of BSA nanoparticles. We found a threshold dose of 20,000 nanoparticles per macrophage uptake in vitro and a saturation dose of 0.3 trillion nanoparticles in tumor-bearing mice. In vivo efficacy and safety evaluations demonstrated that the threshold doses of blank BSA nanoparticles could significantly improve the efficacy and safety of ABP against tumors compared with ABP alone. In this study, the delivery efficiency of ABP was improved by using blank nanoparticles to saturate Kupffer cells, which provided a new approach to studying the Kupffer cell saturation threshold and thus a new scheme for improving the curative effect of ABP.

Metabolite interactions in prostatic fluid mimics assessed by 1H NMR.

INTRODUCTION: Molecular interactions in prostatic fluid are of biological interest and may affect MRI and MRS of the prostate. We investigated the existence of interactions between the major components of this fluid: spermine, citrate and myoinositol, metal ions, including zinc, and proteins. MATERIALS AND METHODS: Solutions of 90 mM citrate, 18 mM spermine and 6 mM myo-inositol, mimicking expressed prostatic fluid, were investigated by 1H NMR using changes in T2 relaxation and chemical shift as markers for interactions. RESULTS AND DISCUSSION: Adding to this metabolite mixture the ions Na+ , K+, Ca++, Mg++ and Zn++, decreased the T2 relaxation times of citrate and spermine protons by factors of 3 and 2, respectively, with Zn++ causing the largest effect, indicating ion-metabolite interactions. The T2 of 18 mM spermine dropped by a factor of 2 upon addition with 90 mM citrate, but no effect on T2 was seen with myo-inositol pointing to a specific citrate-spermine interaction. Moreover, the T2 of citrate in the presence of spermine decreased by adding metal ions and increasing amounts of Zn++, indicating complexation of citrate and spermine with metal ions, particularly with Zn. The addition of bovine serum albumin (BSA), as an index protein, substantially further decreased the T2 of spermine and citrate implying the formation of a transient spermine-metal ion-citrate-BSA complex. Finally, we found that the T2 of citrate in extracellular fluid of prostate cancer cells, as a mimic of fluid in cancerous prostates, decreased by adding fetal calf serum, indicating protein binding.

Albumin Binds Doxorubicin via Self-Assembling Dyes as Specific Polymolecular Ligands.

Congo red (CR) type self-assembled ribbon-like structures (SRLS) were previously shown to interact with some proteins, including albumin. SRLS also complex with some drugs with a flat, ring-shaped structure with aromatic characteristics, intercalating them into their ribbon structure. The combination of interaction with proteins and drug binding by SRLS enables the use of such systems for immunotargeting. It is especially interesting in the case of chemotherapeutic agents. The present experiments aimed to show that the model carrier system composed of supramolecular albumin and Congo red efficiently binds doxorubicin (Dox) and that the drug can be released at reduced pH. The presented results come from the studies on such complexes differing in the molar ratio of CR to Dox. The following methods were used for the analysis: electrophoresis, dialysis, gel filtration, spectral analysis, and analysis of the size of the hydrodynamic radius using the dynamic light scattering method (DLS). The applied methods confirmed the formation of the CR-Dox complex, with large dimensions and changed properties compared with free CR. The presented results show that albumin binds both CR and its complex with Dox. Various CR-Dox molar ratios, 5:1, 2:1, and 1:1, were analyzed. The confirmation of the possibility of releasing the drug from the carriers thus formed was also obtained. The presented research is important due to the search for optimal solutions for the use of SRLS in drug immunotargeting, with particular emphasis on chemotherapeutic agents.

A TCF-Based Carbon Monoxide NIR-Probe without the Interference of BSA and Its Application in Living Cells.

As toxic gaseous pollution, carbon monoxide (CO) plays an essential role in many pathological and physiological processes, well-known as the third gasotransmitter. Owning to the reducibility of CO, the Pd0-mediated Tsuji-Trost reaction has drawn much attention in CO detection in vitro and in vivo, using allyl ester and allyl ether caged fluorophores as probes and PdCl2 as co-probes. Because of its higher decaging reactivity than allyl ether in the Pd0-mediated Tsuji-Trost reaction, the allyl ester group is more popular in CO probe design. However, during the application of allyl ester caged probes, it was found that bovine serum albumin (BSA) in the fetal bovine serum (FBS), an irreplaceable nutrient in cell culture media, could hydrolyze the allyl ester bond, and thus give erroneous imaging results. In this work, dicyanomethylenedihydrofuran (TCF) and dicyanoisophorone (DCI) were selected as electron acceptors for constructing near-infrared-emission fluorophores with electron donor phenolic OH. An allyl ester and allyl ether group were installed onto TCF-OH and DCI-OH, constructing four potential CO fluorescent probes, TCF-ester, TCF-ether, DCI-ester, and DCI-ether. Our data revealed that ester bonds of TCF-ester and DCI-ester could completely hydrolyze in 20 min, but ether bonds in TCF-ether and DCI-ether tolerate the hydrolysis of BSA and no released fluorescence was observed even up to 2 h. Moreover, passing through the screen, it was concluded that TCF-ether is superior to DCI-ether due to its higher reactivity in a Pd0-mediated Tsuji-Trost reaction. Also, the large stokes shift of TCF-OH, absorption and emission at 408 nm and 618 nm respectively, make TCF-ether desirable for fluorescent imaging because of differentiating signals from the excitation light source. Lastly, TCF-ether has been successfully applied to the detection of CO in H9C2 cells.

Synthesis and physicochemical characterization of rhamnolipid fabricated fucoxanthin loaded bovine serum albumin nanoparticles supported by simulation studies.

BACKGROUND: Fucoxanthin is a hydrophobic carotenoid with many beneficial biological activities. However, due to low aqueous solubility their clinical efficacy is limited thus leading to poor oral bioavailability. To address this issue, we encapsulated fucoxanthin in rhamnolipid fabricated bovine serum albumin (BSA) loaded nanoparticles (LNPs) for improving solubility dependent bioavailability of fucoxanthin. RESULTS: These synthesized LNPs were characterized by dynamic light scattering (DLS), ultraviolet (UV)-visible spectrophotometry, high-performance liquid chromatography (HPLC), Fourier-transform infrared (FTIR), scanning electron microscopy (SEM), differential scanning calorimetry (DSC). Our results showed that LNPs were spherical in shape with particle size around 180 nm along with positive zeta potential. The encapsulation efficiency and loading efficiency calculated for LNPs were 69.66 ± 1.5% and 14 ± 0.2%, respectively. The antioxidant assay of LNPs indicate high radical scavenging activity compared to pure fucoxanthin. Besides this, our release studies indicates that drug release occur from the matrix of nanocarrier system through diffusion based on concentration. Thus, these findings indicate successful encapsulation of fucoxanthin, with improved solubility thereby leading to increased bioavailability. This nano formulation is derived from components which are FDA approved that could be exploited for encapsulating other vital nutraceutical molecules. CONCLUSION: Overall, our results showed successful synthesis of biodegradable nanocarrier for delivering fucoxanthin supported by molecular docking, molecular dynamics simulation and thermodynamics of free binding energy studies. © 2022 Society of Chemical Industry.

Using fluorescence and circular dichroism (CD) spectroscopy to investigate the interaction between di-n-butyl phthalate and bovine serum albumin.

The interaction between di-n-butyl phthalate (DBP) and bovine serum albumin (BSA) in physiological Tris-HCl buffer at pH 7.4 was investigated by fluorescence quenching technique. By analyzing the fluorescence spectrum and intensity, it was observed that the DBP had a strong ability to quench the intrinsic fluorescence of BSA through a static quenching procedure. The binding constants K and the number of binding sites n of DBP with BSA were calculated to be 0.11 × 102 L·mol-1 and 0.52 at 298 K, respectively. The thermodynamic parameters of enthalpy change (ΔH) and entropy change (ΔS) were also calculated to be positive showing that hydrophobic forces might play a major role in the binding of DBP to BSA. The binding process was spontaneous in which Gibbs free energy change (ΔG) was negative. The distance (r) between the donor (BSA) and acceptor (DBP) was calculated to be 2.02 nm based on Forster's non-radiative energy transfer theory, which indicated that the energy transfer from BSA to DBP occurs with a high possibility. The synchronous fluorescence, three-dimensional fluorescence, and circular dichroism (CD) spectra showed that the binding of di-n-butyl phthalate to BSA induced conformational changes in BSA. The interaction between DBP and BSA can help researchers better understand the nature of poisons and serve people in the right way with first aid and detoxification.

Endocytosed nanogold fiducials for improved in-situ cryo-electron tomography tilt-series alignment.

Cryo-electron tomography (CET) on cryo-focused ion beam (FIB)-milled lamellae is becoming a powerful technique for determining the structure of macromolecular complexes in their native cellular environment. Prior to tomogram reconstruction, CET tilt-series recorded on FIB lamellae need to be aligned. Traditionally, CET tilt-series alignment is performed with 5-20 nm gold fiducials, but it has thus far proven difficult to apply this to FIB lamellae of eukaryotic cells. In here, we describe a simple method to allow uptake of bovine serum albumin (BSA)-gold fiducials into mammalian cells via endocytosis, which can subsequently be used as fiducials for tilt-series alignment of cryo-FIB lamellae. We compare the alignment of tilt-series with BSA-gold fiducials to fiducial-less patch-tracking, and find better alignment results with BSA-gold. This technique can contribute to understand cells at a structural and ultrastructural level with both cryo- and room-temperature electron tomography. Furthermore, fluorescently labeled BSA-gold has the potential to be used as fiducials for correlative light and electron microscopy studies.

Protein intrinsic viscosity determination with the Viscosizer TD instrument: reaching beyond the initially expected applications.

Intrinsic viscosity is a key hydrodynamic parameter to understand molecular structure and hydration, as well as intramolecular interactions. Commercially available instruments measure intrinsic viscosity by recording the macromolecular mobility in a capillary. These instruments monitor Taylor dispersion using an absorbance or fluorescence detector. By design, these instruments behave like U-tube viscometers. To our knowledge, there are no studies to date showing that the Viscosizer TD instrument (Malvern-Panalytical) is able to measure the intrinsic viscosity of macromolecules. In this study, we then performed our assays on the Poly(ethylene oxide) polymer (PEO), used classically as a standard for viscometry measurements and on three model proteins: the bovine serum albumin (BSA), the bevacizumab monoclonal antibody, and the RTX Repeat Domain (RD) of the adenylate cyclase toxin of Bordetella pertussis (CyaA). The presence of P20 in the samples is critical to get reliable results. The data obtained with our in-house protocol show a strong correlation with intrinsic viscosity values obtained using conventional techniques. However, with respect to them, our measurements could be performed at relatively low concentrations, between 2 and 5 mg/ml, using only 7 µL per injection. Altogether, our results show that the Viscosizer TD instrument is able to measure intrinsic viscosities in a straightforward manner. This simple and innovative approach should give a new boost to intrinsic viscosity measurements and should reignite the interest of biophysicists, immunologists, structural biologists and other researchers for this key physicochemical parameter.

Mineral Scaling on Reverse Osmosis Membranes: Role of Mass, Orientation, and Crystallinity on Permeability.

Prior mineral scaling investigations mainly studied the effects of membrane surface properties rather than on the mineral properties and their impact on membrane permeability. In our study, mass, crystal growth orientation, and crystallinity of mineral precipitates on membranes, as well as their effects on membrane permeability have been investigated. Gypsum scaling tests on bare and bovine serum albumin (BSA)-conditioned membranes were conducted under different saturation indices. Results show that a longer scaling period was required for BSA-conditioned membranes to reach the same membrane permeate flux decline as bare membranes. Though the final reduced permeability was the same for both two membranes, the masses of the mineral precipitates on BSA-conditioned membranes were around two times more than those on bare membranes. Further mineral characterizations confirmed that different permeability decay rates of both types of the membrane were attributed to the differences in growth orientations rather than amounts of gypsum precipitates. Moreover, BSA-conditioned layers with high carboxylic density and specific molecular structure could stabilize bassanite and disrupt the oriented growth to inhibit the formation of needle-like gypsum crystals as observed on bare membranes, thus resulting in lower surface coverage with scales on membranes and alleviating the detrimental scaling effect on membrane permeability.

High-Throughput Analysis of Amino Acids for Protein Quantification in Plant and Animal-Derived Samples Using High Resolution Mass Spectrometry.

Current methods for measuring the abundance of proteogenic amino acids in plants require derivatisation, extended run times, very sensitive pH adjustments of the protein hydrolysates, and the use of buffers in the chromatographic phases. Here, we describe a fast liquid chromatography-mass spectrometry (LC-MS) method for the determination of amino acids that requires only three steps: hydrolysis, neutralisation, and sample dilution with a borate buffer solution for pH and retention time stability. The method shows excellent repeatability (repeated consecutive injections) and reproducibility (repeated hydrolysis) in the amino acid content, peak area, and retention time for all the standard amino acids. The chromatographic run time is 20 min with a reproducibility and repeatability of <1% for the retention time and <11% for the peak area of the BSA and quality control (QC) lentil samples. The reproducibility of the total protein levels in the hydrolysis batches 1-4 was <12% for the BSA and the lentil samples. The level of detection on column was below 0.1 µM for most amino acids (mean 0.017 µM).

Preparation of Antihypertensive Drugs in Biological Matrix with Solvent Front Position Extraction for LC-MS/MS Analysis.

Solvent front position extraction procedure was used to prepare biological samples containing selected antihypertensive drugs (ramipril, lercanidipine, indapamide, valsartan, hydrochlorothiazide, perindopril, and nebivolol). Substances separated from the biological matrix components (bovine serum albumin) were quantified by means of liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Sample preparation process was performed with the use of a prototype horizontal chamber with a moving pipette driven by a 3D printer mechanism enabling a controlled eluent flow velocity. Application of this device was advantageous for simultaneous preparation of several samples for further quantitative analysis, with a synchronized reduction of manual operations and solvent consumption. Quantitative results obtained for the majority of the investigated antihypertensive drugs in a complex biological matrix were satisfactory. The values of the %RSD were around 5% for six of the seven substances (with the exception of indapamide). The method exhibits a suitable accuracy (the relative error percentage was below 10% for most drugs). The values of LOD and LOQ were in the range of 1.19 µg/L-8.53 µg/L and 3.61 µg/L-25.8 µg/L, respectively.

Preparation of Amorphous Solid Dispersions by Cryomilling: Chemical and Physical Concerns Related to Active Pharmaceutical Ingredients and Carriers.

In this work, a chemical (and physical) evaluation of cryogenic milling to manufacture amorphous solid dispersions (ASDs) is provided to support novel mechanistic insights in the cryomilling process. Cryogenic milling devices are considered as reactors in which both physical transitions (reduction in crystallite size, polymorphic transformations, accumulation of crystallite defects, and partial or complete amorphization) and chemical reactions (chemical decomposition, etc.) can be mechanically triggered. In-depth characterization of active pharmaceutical ingredient (API) (content determination) and polymer (viscosity, molecular weight, dynamic vapor sorption, Fourier transform infrared spectroscopy, dynamic light scattering, and ANS and thioflavin T staining) chemical decomposition demonstrated APIs to be more prone to chemical degradation in case of presence of a polymer. A significant reduction of the polymer chain length was observed and in case of BSA denaturation/aggregation. Hence, mechanochemical activation process(es) for amorphization and ASD manufacturing cannot be regarded as a mild technique, as generally put forward, and one needs to be aware of chemical degradation of both APIs and polymers.

Bovine serum albumin affects N-glycoforms of murine IgG monoclonal antibody purified from hybridoma supernatants.

Immunoglobulin G (IgG) is a class of monoclonal antibodies (mAbs) commonly produced in mammalian cell lines. These cell lines are grown in finely adjusted culture media, which contain components that may impact glycoforms. As variation of N-glycoforms can impact the biological properties of IgGs, medium composition should be controlled. Here, we studied the effects on IgG N-glycoforms of different components in hybridoma culture media, specifically compared bovine serum albumin (BSA) with other small molecules, using a matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight multistage mass spectrometry (MALDI-QIT-TOF MSn)-based approach. We show that small molecular additives caused little change in glycan species, though a number of these reagents, especially glutamine, affected levels of glycosylation. In comparison, the addition of macromolecular protein BSA significantly changed IgG N-glycan patterns, not only in species but also in glycosylation levels. Together, our finding suggests that BSA increases the complexity of IgG N-glycoforms, thus raising the difficulty in maintaining glycoforms consistency during antibody production. Therefore, the effect of BSA on IgG N-glycans should be considered when designing optimal medium formulations for IgG production. KEY POINTS: • Small molecular medium additives only affect glycosylation levels of IgG N-glycans. • BSA significantly changes IgG N-glycoforms as a medium additive. • BSA's skewing of IgG N-glycoforms should be considered in IgG production.

Investigation on the interaction of food colorant Sudan III with bovine serum albumin using spectroscopic and molecular docking methods.

Sudan III is a coloring agent used in chemical industries and food additives. This article uses spectroscopic and molecular docking methods to investigate the interaction of Sudan III with bovine serum albumin (BSA) under a physiological condition. Spectroscopic analysis of the emission quenching revealed that the quenching mechanism of BSA by Sudan III was static. The binding sites and constants of Sudan III-BSA complex were observed to be from 0.72 and 6.41 × 102 L·mol-1 to 0.69 and 5.83 × 102 L·mol-1 at 298 and 310 K, respectively. The enthalpy change (ΔH) and entropy change (ΔS) revealed that van der Waals forces and hydrogen bonds stabilized the Sudan III-BSA complex. Energy transfer from tryptophan to Sudan III occurred by a fluorescence resonance energy transfer mechanism, and the r distance (3.32 nm) had been determined. The results of UV-Vis absorption, synchronous, three-dimensional fluorescence, and circular dichroism spectra showed that Sudan III induced conformational changes of BSA. Molecular docking studies revealed that Sudan III situated within subdomain IIA of BSA. A study on the interaction between Sudan III and BSA was of fundamental importance for providing more information about the potential toxicological effect of chemicals at the molecular level.