Bovine Serum Albumin Complex Research Articles

Investigations on the complexation and binding mechanism of bovine serum albumin with Ag-doped TiO2 nanoparticles.

It is essential to study the interactions between nanoparticles and proteins to better understand the biological interactions of nanoparticles. In this study, we studied the protein adsorption mode on the surface of Ag-doped TiO2 nanoparticles (NPs) using a model protein, bovine serum albumin (BSA). The mechanism of binding BSA to the Ag-doped TiO2 NPs was studied by applying fluorescence quenching, absorbance measurements, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy techniques. The strong binding between BSA and Ag-doped TiO2 NPs was confirmed by a high value of binding constant (K = 2.65 × 105 L mol-1). We also studied the thermal stability of BSA in the presence of the Ag-doped TiO2 NPs. Thermodynamic parameters indicated that the adsorption of BSA on the Ag-doped TiO2 NPs was a spontaneous, natural and exothermic process. The effect of Ag-doped TiO2 NPs on the transportation function of BSA was also studied using a fluorescence spectroscopic technique. Fluorescence spectroscopic data suggested the existence of a strong interaction between BSA and the surface of the Ag-doped TiO2 NPs, which indicated that the binding affinities of some selected amino acids in BSA changed. This, in turn, clearly confirms that the Ag-doped TiO2 NPs affect the transportation capability of BSA in blood.

Investigation of the molecular interaction between apraclonidine, an α2-adrenergic receptor agonist, and bovine serum albumin using fluorescence and molecular docking techniques

Apraclonidine (APR) is a potent and selective α2-adrenergic receptor agonist used in the diagnosis of Horner’s Syndrome, and the residuals of APR that accumulate in tissues of animals can cause central nervous and cardiovascular systems influences in humans. Therefore, to understand the influence of APR on human health, we examined the interaction of APR with the carrier protein in plasma, bovine serum albumin (BSA). The BSA fluorescence signal was quenched due to the APU–BSA complex formation and a weak binding affinity was estimated between APR and BSA. The inclusion of fluorescence, UV–vis absorption, molecular docking, and dynamics simulation techniques employed to broadly investigate the combination of APR with BSA at typical physiological conditions. The thermodynamic results revealed that enthalpy (ΔH0) and entropy (ΔS0) changes were computed as +11.14 kJ mol−1 and +97.56 J mol−1 K−1, respectively, which represented the binding is principally entropy-driven and the hydrophobic forces acting a significant role in the reaction. Analysis of synchronous and 3-D fluorescence signals revealed microenvironmental variations close to BSA’s Trp and Tyr residues upon APR addition. Both the competitive site marker as well as molecular docking results detected that APR exhibited a stronger binding affinity towards Drug Site 2 (DS2) compared to Drug Site 1 (DS1).

ICAM‐1‐Expressing Extracellular Vesicles Enable Sequential Delivery Barrier‐Breaking and Microenvironment Modulating for Glioblastoma Stem Cell Inhibition

AbstractGlioma stem‐like cell (GSC) inhibition is a promising strategy for glioblastoma therapy. However, it is hampered by two main obstacles: first, delivery barriers, including the blood–brain barrier (BBB) and deep tumor‐penetrating obstacles, hinder sufficient drug accumulation in GSCs that often reside in hypoxic niches deep in glioblastoma; second, hypoxia and the immunosuppressive tumor microenvironment (TME) limit the therapeutic responses of GSC inhibition strategies. An intercellular cell adhesion molecule 1 (ICAM‐1)‐expressing M1 macrophage‐derived small extracellular vesicles loaded with a complex of bovine serum albumin, MnO2, and the small‐molecule inhibitor PTC209 (IsEV‐BMP) is constructed to achieve sequential barrier‐breaking delivery and simultaneous TME modulation to boost GSC inhibition. ICAM‐1 facilitates endothelial transcytosis‐mediated BBB crossing, microglial transcytosis‐mediated deep tumor penetration, and internalization of IsEV‐BMPs in GSCs. Hypoxia alleviation inhibits the binding of hypoxia‐inducible factor 1α (HIF‐1α) to hypoxia‐responsive elements on the gene promoters of cancer stem cell‐related markers and CD47 in GSCs, thereby improving PTC209 sensitivity and immune phagocytosis of GSCs. IsEV‐BMP achieves simultaneous microglial polarity transition and immuno‐microenvironmental modulation during microglial transcytosis‐mediated deep tumor penetration. This study provides a strategy to overcome the sequential delivery barrier for effective glioblastoma treatment.

Adducts of bovine serum albumin and binuclear nitrosyl iron complex with thiosulfate ligands: a molecular docking and quantum chemical study

Adducts of bovine serum albumin and binuclear nitrosyl iron complex with thiosulfate ligands: a molecular docking and quantum chemical study

A Novel Pickering High Internal Phase Emulsion Gels Fabricated by Starch Nanocrystals and Bovine Serum Albumin Complexes

A Novel Pickering High Internal Phase Emulsion Gels Fabricated by Starch Nanocrystals and Bovine Serum Albumin Complexes

Interactions of three berberine mid-chain fatty acid salts with bovine serum albumin (BSA): Spectroscopic analysis and molecular docking

In this paper, the interaction of three berberine mid-chain fatty acid salts ([BBR][FAs]), viz. berberine caproate ([BBR][CAP]), berberine heptylate ([BBR][HEP]) and berberine octoate ([BBR][OCT]), with bovine serum albumin (BSA) was studied by means of UV–visible absorption spectroscopy, fluorescence spectroscopy, fourier transform infrared spectroscopy (FT-IR) and molecular docking techniques. Fluorescence experiments revealed that three berberine salts quench the fluorescence of BSA by static quenching mechanism resulted from a stable [BBR][FAs]-BSA complex formation. The stoichiometric numbers of [BBR][FAs]-BSA complexes were found to be 1:1. Synchronous and three-dimensional fluorescence spectra as well as FT-IR demonstrated that the binding of [BBR][FAs] altered the microenvironment and conformation of BSA. The binding average distance from [BBR][FAs] to BSA (3.2–3.5 nm) was determined according to Förster energy transfer theory. Site probe investigation showed that [BBR][FAs] bound to BSA active site I (sub-domain IIA). The binding promotes the esterase-like activity of BSA. The molecular docking results confirmed the fluorescence competition findings and provided the type of binding forces. Furthermore, the relationship between the anionic chain length of [BBR][FAs] and the interaction was explored, and the positive correlation was found.

Fluorinated Chitosan‐Mediated Transepithelial Delivery of Intravesical Dual‐Drug Immunotherapeutic for Bladder Cancer Therapy

AbstractIntravesical instillation‐based immunotherapy for bladder‐preservation of bladder cancer (BCa) treatment has not been explored. In the current study, two irrigated nano‐formulations of immunological adjuvant‐fluorinated chitosan (FCS) and therapeutic antibody‐FCS are developed for synergistic immunotherapy against BCa. In this system, FCS is employed as a transepithelial carrier to assemble with bovine serum albumin (BSA)‐flubendazole (FBZ) complex and anti‐PD‐1 (αPD‐1) respectively, to facilitate efficient transepithelial delivery of intravesical FBZ‐BSA and αPD‐1 though transiently regulating the distribution of tight junction proteins on bladder epithelium. In addition, the FBZ–BSA complex exhibits tumor microenvironment‐responsive charge reversal and BSA carrier‐broken effects to drive tumor‐targeted dissociation and drug release of FBZ–BSA/FCS. Moreover, FBZ not only promotes apoptosis and inhibits glycolysis in cancer cells but also displays adjuvant properties to activate potent immune responses through IL‐12/IFN‐γ pathway. Interestingly, combined perfusion of FBZ–BSA/FCS and αPD‐1/FCS nano‐formulations can significantly activate the tumor immune microenvironment, especially CD8+ T cell infiltration and αPD‐1 sensitivity, and finally remarkably inhibit tumor growth in the mouse orthotopic BCa model. Thus, this study presents a novel intravesical delivery platform for ICB antibodies and a smart adjuvant system to achieve potent synergy immunotherapy, which is promising for bladder‐preserving treatment against BCa.

Nanofibrous polyelectrolyte complex incorporated BSA-alginate composite bioink for 3D bioprinting of bone mimicking constructs

Although several bioinks have been developed for 3D bioprinting applications, the lack of optimal printability, mechanical properties, and adequate cell response has limited their practical applicability. Therefore, this work reports the development of a composite bioink consisting of bovine serum albumin (BSA), alginate, and self-assembled nanofibrous polyelectrolyte complex aggregates of gelatin and chitosan (PEC-GC). The nanofibrous PEC-GC aggregates were prepared and incorporated into the bioink in varying concentrations (0 % to 3 %). The bioink samples were bioprinted and crosslinked post-printing by calcium chloride. The average nanofiber diameter of PEC-GC was 62 ± 15 nm. It was demonstrated that PEC-GC improves the printability and cellular adhesion of the developed bioink and modulates the swelling ratio, degradation rate, and mechanical properties of the fabricated scaffold. The in vitro results revealed that the bioink with 2 % PEC-GC had the best post-printing cell viability of the encapsulated MG63 osteosarcoma cells and well oragnized stress fibers, indicating enhanced cell adhesion. The cell viability was >90 %, as observed from the MTT assay. The composite bioink also showed osteogenic potential, as confirmed by the estimation of alkaline phosphatase activity and collagen synthesis assay. This study successfully fabricated a high-shape fidelity bioink with potential in bone tissue engineering.

Dual quenching nature of bovine serum albumin and dopamine complex revealed using multi-spectral and docking studies

In the present study, dopamine interaction with bovine serum albumin was investigated using fluorescence, UV–vis, Raman, CV, FTIR and molecular modeling techniques. The fluorescence study of the quenching was carried out at an excitation wavelength of 296.0 nm, and the emission was measured in the range of 300–600 nm. BSA was quenched through both static and dynamic mechanisms. Thermodynamic parameters were derived to study the binding forces involved. The average binding distance between BSA and dopamine was determined using FÖrsters theory. Lifetime measurements, synchronous fluorescence and 3D spectra supported the hypotheses derived from the fluorescence data. The presence of some common co-existent drugs and the effect of essential biological metal ions were also investigated.

Insight into the interaction of isochroman with bovine serum albumin: extensive experimental and computational investigations

The way therapeutic compounds interact with serum protein provides valuable information on their pharmacokinetics, toxicity, effectiveness, and even their structural-related information. Isochroman (IC) is a phytochemical compound obtained from the leaves of Olea europea plant. The derivatives of IC have various pharmacological properties including antidepressants, antihistamines, antiinflammation, anticonvulsants, appetite depressants, etc. The binding of small molecules to bovine serum albumin (BSA) is useful to ensure their efficacy. Thus, in this study, we have found out the binding mode of IC with BSA using several spectroscopic and in silico studies. UV and fluorescence spectroscopy suggested the complex formation between IC and BSA with a binding constant of 103 M−1. IC resulted in fluorescence quenching in BSA through static mechanism. The microenvironmental and conformational changes in BSA were confirmed using synchronous and three-dimensional studies. Site marker experiment revealed the IC binding in site-III of BSA. The influence of vitamins, metals and β-cyclodextrin (β-CD) on binding constant of IC–BSA complex was also examined. Circular dichroism spectra showed that α-helical of BSA decreased upon interaction with IC. Computational and experimental results were complimentary with one another and assisted in determining the binding sites, nature of bonds and amino acids included in the IC–BSA complex formation. Communicated by Ramaswamy H. Sarma

Exploration of the intermolecular isoproturon–bovine serum albumin combination: Biophysical and computational prospects

Isoproturon (IPU), a selective systemic herbicide that is frequently applied to eradicate large-leaved weeds and annual grasses in agricultural fields. However, the accumulation of IPU residues during food production might negatively affect on human and animal health. To comprehend the impact of IPU on the mankind and environment, the interaction analysis of IPU with plasma protein is crucial. This work used a variety of techniques, including fluorescence, UV–vis absorption, computational docking and dynamics simulation to extensively examine the combination of IPU with bovine serum albumin (BSA) under typical physiological settings. IPU quenched the BSA fluorescence and form the IPU–BSA complex through a static process. A weak binding affinity was anticipated (binding constant = 4.68–2.48 × 103 M−1) for IPU–BSA interaction. Thermodynamic results revealed that the IPU–BSA binding reaction was a spontaneous process, predominantly governed by hydrogen bonds, van der Waals and hydrophobic forces, which was well supported by molecular docking analysis. Synchronous and three-dimensional fluorescence spectral results exhibited microenvironmental fluctuations near the protein’s Trp and Tyr residues with added IPU, but inclusion of IPU to BSA was observed to enhance the protein’s thermal constancy. Site marker displacement results demonstrated that IPU desirably binds to BSA at site II. An in silico findings also unveiled the primary binding arrangement of IPU was placed at site II of BSA and surrounded by hydrophobic and polar residues. Molecular dynamics simulation results suggested the IPU–BSA complex persisted compact and stable.

Magnetically navigated microbubbles coated with albumin/polyarginine and superparamagnetic iron oxide nanoparticles

While microbubbles (MB) are routinely used for ultrasound (US) imaging, magnetic MB are increasingly explored as they can be guided to specific sites of interest by applied magnetic field gradient. This requires the MB shell composition tuning to prolong MB stability and provide functionalization capabilities with magnetic nanoparticles. Hence, we developed air-filled MB stabilized by a protein–polymer complex of bovine serum albumin (BSA) and poly-L-arginine (pArg) of different molecular weights, showing that pArg of moderate molecular weight distribution (15–70 kDa) enabled MB with greater stability and acoustic response while preserving MB narrow diameters and the relative viability of THP-1 cells after 48 h of incubation. After MB functionalization with superparamagnetic iron oxide nanoparticles (SPION), magnetic moment values provided by single MB confirmed the sufficient SPION deposition onto BSA + pArg MB shells. During MB magnetic navigation in a blood vessel mimicking phantom with magnetic tweezers and in a Petri dish with adherent mouse renal carcinoma cell line, we demonstrated the effectiveness of magnetic MB localization in the desired area by magnetic field gradient. Magnetic MB co-localization with cells was further exploited for effective doxorubicin delivery with drug-loaded MB. Taken together, these findings open new avenues in control over albumin MB properties and magnetic navigation of SPION-loaded MB, which can envisage their applications in diagnostic and therapeutic needs.

Crystalline silica-induced proinflammatory eicosanoid storm in novel alveolar macrophage model quelled by docosahexaenoic acid supplementation.

Phagocytosis of inhaled crystalline silica (cSiO2) particles by tissue-resident alveolar macrophages (AMs) initiates generation of proinflammatory eicosanoids derived from the ω-6 polyunsaturated fatty acid (PUFA) arachidonic acid (ARA) that contribute to chronic inflammatory disease in the lung. While supplementation with the ω-3 PUFA docosahexaenoic acid (DHA) may influence injurious cSiO2-triggered oxylipin responses, in vitro investigation of this hypothesis in physiologically relevant AMs is challenging due to their short-lived nature and low recovery numbers from mouse lungs. To overcome these challenges, we employed fetal liver-derived alveolar-like macrophages (FLAMs), a self-renewing surrogate that is phenotypically representative of primary lung AMs, to discern how DHA influences cSiO2-induced eicosanoids. We first compared how delivery of 25 µM DHA as ethanolic suspensions or as bovine serum albumin (BSA) complexes to C57BL/6 FLAMs impacts phospholipid fatty acid content. We subsequently treated FLAMs with 25 µM ethanolic DHA or ethanol vehicle (VEH) for 24h, with or without LPS priming for 2h, and with or without cSiO2 for 1.5 or 4h and then measured oxylipin production by LC-MS lipidomics targeting for 156 oxylipins. Results were further related to concurrent proinflammatory cytokine production and cell death induction. DHA delivery as ethanolic suspensions or BSA complexes were similarly effective at increasing ω-3 PUFA content of phospholipids while decreasing the ω-6 PUFA arachidonic acid (ARA) and the ω-9 monounsaturated fatty acid oleic acid. cSiO2 time-dependently elicited myriad ARA-derived eicosanoids consisting of prostaglandins, leukotrienes, thromboxanes, and hydroxyeicosatetraenoic acids in unprimed and LPS-primed FLAMs. This cSiO2-induced eicosanoid storm was dramatically suppressed in DHA-supplemented FLAMs which instead produced potentially pro-resolving DHA-derived docosanoids. cSiO2 elicited marked IL-1α, IL-1β, and TNF-α release after 1.5 and 4h of cSiO2 exposure in LPS-primed FLAMs which was significantly inhibited by DHA. DHA did not affect cSiO2-triggered death induction in unprimed FLAMs but modestly enhanced it in LPS-primed FLAMs. FLAMs are amenable to lipidome modulation by DHA which suppresses cSiO2-triggered production of ARA-derived eicosanoids and proinflammatory cytokines. FLAMs are a potential in vitro alternative to primary AMs for investigating interventions against early toxicant-triggered inflammation in the lung.

Bridging Molecular Docking to Molecular Dynamics to Enlighten Recognition Processes of Tailored D-A/D-A-D Types' AIEgens with HSA/BSA

Fluorescence imaging-assisted photodynamic therapy (PDT) allows accurate tumor visualization and potentially prevents long-term side effects of cancer. Therefore, the development of photosensitizers emitting light, particularly in the near-infrared region (NIR), is essential for enhancing the efficacy of cancer therapy. On this premise, the formation of a stabilized complex between an organic dye and a target macromolecule improves fluorescence efficiency. In this scope, we performed a detailed molecular dock-ing study of Donor (D)-Acceptor (A) or D-A-D type luminogens with two blood proteins; namely bovine serum albumin (BSA) and human serum albumin (HSA), which appeared as robust carriers of several pharmaceuticals against preliminary cancer diseases. Our results revealed that the binding scores of the Dn-An or Dn-An-Dn:BSA complexes ranged from -8.5 to -11.7 kcal/mol while Dn-An or Dn-An-Dn:HSA complexes showed scores varying from -8.4 to -10.5 kcal/mol. Subsequently, molecular dynamics simu-lations were also performed for the best-docked ligands: macromolecule complexes; namely D1A1D1:BSA and D1A1:HSA, to enlighten various structural parameters. Based on the predicted root-mean-square deviation (RMSD) values (on average), the D1A1D1:BSA complex was found to be 0.319 nm, while the D1A1:HSA complex was determined as 0.284 nm. In addition, the root-mean-square fluctuations (RMSF) analyses (on average) revealed that D1A1D1:BSA (0.152 nm) was slightly more flexible than D1A1:HSA (0.160 nm).

Simple and sensitive fluorescence detection of trypsin with Cu2+-Bovine serum albumin complex as a peroxidase mimic

Trypsin is a serine protease playing a key role in regulating pancreatic exocrine function and can be applied as a marker for the diagnosis of pancreatitis. In this work, a convenient and sensitive fluorescent assay was developed toward trypsin. Hydrogen peroxide slowly oxidized a non-fluorescent o-phenylenediamine (OPD) into a fluorescent product 2,3-diaminophenothiazine (DAP) under the catalytic from copper ions. After the introduction of bovine serum albumin (BSA), the combination of BSA with copper ions formed a peroxidase mimic and significantly accelerated the reaction rate. As an efficient protease, trypsin cleaved the lysine and arginine residues in BSA. This destroyed the binding between Cu2+ and BSA, and brought in a reduction of the catalytic effect. The accompanying decrease in fluorescence provided a response to trypsin in the range of 0.01–600 ng/mL, with a detection limit of 0.007 ng/mL. The scheme had a good selectivity and was successfully applied to the detection of real samples.

Spectroscopic aspects on the interaction of nisin with serum albumin: thermodynamic and kinetic studies.

Nisin is a bacteriocin produced by Streptococcus and Lactococcus species and has antimicrobial activity against other bacteria. Nisin omits the need to use chemical preservatives in food due to its biological preserving properties. In the present in vitro study, we investigated nisin interaction with bovine serum albumin (BSA) using fluorescence spectroscopy and surface plasmon resonance (SPR) analysis to obtain information about the mechanisms of BSA complex formation with nisin. The BSA fluorescence intensity values gradually diminished with rising nisin concentration. The BSA fluorescence quenching analysis indicated that a combined quenching mechanism plays the main role. Finally, the Kb values were reduced with increasing temperature, which is demonstrative of nisin-BSA complex stability decrease at high temperatures. The negative values of ΔH° and ΔS° showed that hydrogen bonds and van der Waals forces are the foremost binding force between BSA and nisin. Meanwhile, the negative values of ΔG° demonstrated the exothermic and random nature of the reaction process. The results of the SPR verified the gained results through the fluorescence spectroscopy investigation, which denoted that the BSA affinity to nisin diminished upon increasing temperature. Overall, fluorescence spectroscopy and SPR results showed that the BSA interaction with nisin decreased with rising temperatures.

Mechanisms of the interaction of bovine serum albumin with quercetin

The peculiarities of the complexation of bovine serum albumin (BSA) and quercetin (Q) are shown in the article. The static mechanism of the formation of BSA-Q conjugates with the predominant effect of hydrogen bonds and hydrophobic interaction is established from fluorescence and absorption spectral data of aqueous solutions at different temperatures. The detailed analysis of an interaction of the molecules is performed in a molecular modeling procedure, including docking and molecular dynamics study. The computation results match the experimental outcome.

Effect of Soft Preheating of Bovine Serum Albumin on the Complexation with Oligochitosan: Structure and Conformation of BSA in the Complex.

Phase analysis, spectroscopic and light scattering methods are applied to investigate the peculiarities of the interaction of oligochitosan (OCHI) with native and preheated BSA as well as the conformational and structural changes of BSA in BSA/OCHI complex. As shown, untreated BSA binds with OCHI mainly forming soluble electrostatic nanocomplexes, with the binding causing an increase in BSA helicity without a change in the local tertiary structure and thermal stability of BSA. In contrast, soft preheating at 56°C enhances the complexation of BSA with OCHI and slightly destabilizes the secondary and local tertiary structures of BSA within the complex particles. Preheating at 64°C (below the irreversible stage of BSA thermodenaturation) leads to further enhancement in the complexation and formation of insoluble complexes stabilized by both Coulomb forces and hydrophobic interactions. The finding can be promising for the preparation of biodegradable BSA/chitosan-based drug delivery systems. This article is protected by copyright. All rights reserved.

In vitro interactions of two pesticides, propazine and quinoxyfen with bovine serum albumin: Spectrofluorometric and molecular docking investigations

Binding mechanisms of two selected pesticides, propazine (PRO) and quinoxyfen (QUI) with bovine serum albumin (BSA) was examined using fluorescence, absorption and molecular docking methods. Intrinsic fluorescence of BSA was quenched in the presence of both PRO and QUI. The quenching was ascertained to be conversely linked to temperature, which suggested the contribution of static quenching process in the PRO–BSA and QUI–BSA complex formations. This results were validated by the enhancement in absorption spectrum of BSA upon binding with PRO and QUI. Binding constant values (Kf = 9.55–0.60 × 10–3 M−1 for PRO–BSA system; Kf = 7.08–5.01 × 102 M−1 for QUI–BSA system) and number of binding site (n) values for the PRO–BSA and QUI–BSA systems at different temperatures affirmed a weak binding strength with a set of equivalent binding sites on BSA. Thermodynamic data obtained for both the PRO–BSA and QUI–BSA interactions predicted that the association process was spontaneous and non-covalent contacts such as hydrophobic interactions, van der Waals forces and hydrogen bonds participated in the binding reactions. This result was further supported by the molecular docking assessments. Three-dimensional spectral results revealed the microenvironmental alterations near tryptophan (Trp) and tyrosine (Tyr) residues in BSA by the addition of PRO and QUI. The docking analysis demonstrated the binding pattern for the PRO–BSA and QUI–BSA systems and disclosed the preferred binding site of both PRO and QUI as site I (subdomain IIA) of BSA.

Binding characteristics of Bruton’s tyrosine kinase inhibitor ibrutinib with bovine serum albumin: multi-spectroscopic combined with molecular simulation

Ibrutinib, as an oral Bruton’s tyrosine kinase inhibitor, has highly potent, covalent irreversible, and other characteristics. In this work, the binding characteristics of ibrutinib with bovine serum albumin were investigated using multi-spectroscopic techniques and molecular simulations. The findings demonstrated that ibrutinib could quench the endogenous fluorescence of bovine serum albumin by static quenching mode, meanwhile bovine serum albumin could also quench the fluorescence of ibrutinib. In the binding process of ibrutinib with bovine serum albumin, ibrutinib caused a bathochromic shift in the absorption band of bovine serum albumin, further indicating that ibrutinib and bovine serum albumin formed a ground state ibrutinib and bovine serum albumin complex. It is confirmed that ibrutinib has a strong affinity to bovine serum albumin due to the binding constant of more 104 M−1. Experimental findings showed that ibrutinib resulted in the decrease in the hydrophobicity or the enhancement in the polarity of the surroundings around Tyr and Trp residues of bovine serum albumin and diminishing in α-helix and β-sheet content of bovine serum albumin. The findings from site-competition experiments confirmed that the binding site of ibrutinib onto bovine serum albumin was the same as that of ibuprofen, that is, ibrutinib bound to the site II' site of bovine serum albumin, which was verified by molecular docking. The findings from thermodynamic analysis revealed that ibrutinib spontaneously bound onto bovine serum albumin through an enthalpy-driven and the driving forces included hydrogen bonding and van der Waals forces being the dominating forces. The findings from molecular dynamics simulation confirmed that some residues such as ARG-208, ALA-209, ALA-212, LEU-326, and LYS-350 made major contributions in ibrutinib and bovine serum albumin complexation.