Adsorption Of Lysozyme Research Articles

Protein adsorption to poly(tetrafluoroethylene) membranes modified with grafted poly(acrylic acid) chains.

Protein adsorption to biomaterial surfaces is important for the function of such materials with anchorage-dependent cell adhesion requiring the presence of adsorbed proteins. The current study evaluated five solid surfaces with poly(acrylic acid) (PAA) grafted from the surface of a poly(tetrafluoroethylene) membrane with respect to the adsorption of serum albumin (SA), lactoferrin (Lf), and lysozyme (Lys) from a phosphate buffer and NaCl solution or water for specific combinations. With the use of x-ray photoelectron spectroscopy, the relative amounts and protein layer thickness were evaluated. SA adsorption was governed by ionic repulsive forces and hydrophobic interactions as evidenced from an increase in the protein adsorption at lower pH (6.5 compared to 7.4) and a correlation with surface coverage when water (pH 6.5) was used as the medium. The adsorption of Lf and Lys followed similar trends for all samples. In general, ionic attractive forces dominated and a strong correlation of increasing protein adsorption with the PAA chain length was evident. This study concluded that all surfaces appear suitable for use in biomaterial applications where tissue ingrowth is desired and that the enhanced protein adsorption in a medium with high ionic strength (e.g., biological fluid) correlates with the PAA chain length rather than the surface coverage.

Carboxylated Cellulose Nanocrystals Developed by Cu-Assisted H2O2 Oxidation as Green Nanocarriers for Efficient Lysozyme Immobilization.

Cellulose nanocrystals (CNCs), having a high specific surface area and versatile surface chemistry, provide considerable potential to interact by various mechanisms with enzymes for nano-immobilization purposes. However, engineering chemically safe CNCs, suitable for edible administrations, presents a significant challenge. A reliable carboxylate form of H-CNCs was formed using H2O2 oxidation of softwood pulp under mild thermal conditions. Negatively charged carboxyl groups (∼0.9 mmol g-1) played a key role in lysozyme immobilization via electrostatic interactions and covalent linkages, as evidenced by Fourier transform infrared and 13C cross-polarization magic angle spinning nuclear magnetic resonance spectroscopies. Adsorption isotherms showed a high loading capacity of H-CNCs (∼240 mg g-1), and fitting the data to the Langmuir model confirmed monolayer coverage of lysozyme on their surface. Using a non-toxic coupling agent, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, lysozyme-conjugated H-CNCs were developed with an immobilization yield of ∼65% and relative catalytic activity of ∼60%, similar to lysozyme adsorption on H-CNCs. These H-CNC-lysozyme nanohybrids, rationally processed via safe and green strategies, are specifically exploitable as catalytically active emulsifiers in food and pharmaceutical sectors.

Microcalorimetric Analysis of the Adsorption of Lysozyme and Cytochrome c onto Cation-Exchange Chromatography Resins: Influence of Temperature on Retention.

We studied the adsorption mechanism of two basic proteins, equine cytochrome c (Cyt) and chicken egg-white lysozyme (Lys), adsorbing onto negatively charged chromatography surfaces. In liquid chromatography, the retention volume of Lys was larger than that of Cyt on negatively charged ion-exchange resins. When the temperature increased, the retention volume of Cyt increased, whereas that of Lys clearly decreased. Both Lys and Cyt share similar physical characteristics, so the opposite behavior with increasing temperatures was surprising, indicating a more complex mechanism of adsorption may be involved. We analyzed the adsorption of these proteins by using isothermal titration calorimetry (ITC). The change in adsorption enthalpy determined by ITC allowed the understanding of the reason for and underlying driving forces of protein adsorption that resulted in this opposite behavior. Large exothermic enthalpies of adsorption were observed for Lys (-43.95 kJ/mol), and Lys adsorption was found to be enthalpically driven. On the other hand, endothermic enthalpies were dominant for Cyt adsorption (32.41 kJ/mol), which was entropically driven. These results indicate that dehydration and release of counterions play a more important role in Cyt adsorption and ionic interaction and hydrogen bridges are more significant in Lys adsorption. Understanding of the adsorption mechanism of proteins onto chromatography resins is essential for modeling and developing new, efficient chromatographic processes.

Computational Investigation of Antifouling Property of Polyacrylamide Brushes.

Antifouling materials and coatings have broad fundamental and practical applications. Strong hydration at polymer surfaces has been proven to be responsible for their antifouling property, but molecular details of interfacial water behaviors and their functional roles in protein resistance remain elusive. Here, we computationally studied the packing structure, surface hydration, and protein resistance of four poly(N-hydroxyalkyl acrylamide) (PAMs) brushes with different carbon spacer lengths (CSLs) using a combination of molecular mechanics (MM), Monte Carlo (MC), and molecular dynamics (MD) simulations. The packing structure of different PAM brushes were first determined and served as a structural basis for further exploring the CSL-dependent dynamics and structure of water molecules on PAM brushes and their surface resistance ability to lysozyme protein. Upon determining an optimal packing structure of PAMs by MM and optimal protein orientation on PAMs by MC, MD simulations further revealed that poly(N-hydroxymethyl acrylamide) (pHMAA), poly(N-(2-hydroxyethyl)acrylamide) (pHEAA), and poly(N-(3-hydroxypropyl)acrylamide) (pHPAA) brushes with shorter CSLs = 1-3 possessed a much stronger binding ability to more water molecules than a poly(N-(5-hydroxypentyl)acrylamide) (pHPenAA) brush with CSL = 5. Consequently, CSL-induced strong surface hydration on pHMAA, pHEAA, and pHPAA brushes led to high surface resistance to lysozyme adsorption, in sharp contrast to lysozyme adsorption on the pHPenAA brush. Computational studies confirmed the experimental results of surface wettability and protein adsorption from surface plasmon resonance, contact angle, and sum frequency generation vibrational spectroscopy, highlighting that small structural variation of CSLs can greatly impact surface hydration and antifouling characteristics of antifouling surfaces, which may provide structural-based design guidelines for new and effective antifouling materials and surfaces.

Protein Adsorption and Bacterial Adhesion Resistance of Cross‐linked Copolymer Hydrogels Based on Poly(2‐methacryloyloxyethyl phosphorylcholine) and Poly(2‐hydroxyethyl methacrylate)

Herein, the preparation of the cross‐linked copolymer hydrogels composed of various weight ratios of 2‐methacryloyloxyethyl phosphorylcholine (MPC) and 2‐hydroxyethyl methacrylate (HEMA) monomers employing free‐radical polymerization was reported. The integration of the MPC monomer into HEMA‐based hydrogels showed good transmittance (>90%) and enhanced the water content retention (79–202%) relative to those of pure HEMA‐based hydrogel. Notably, the MPC‐containing hydrogels (MPC‐H) exhibited the improved anti‐biofouling properties due to the formation of a compact hydration layer produced by the biomimetic MPC units at the hydrogel surface. MPC‐H exhibited a decrease in the bovine serum albumin (BSA) and lysozyme adsorption of approximately 40–70% and 44–65%, respectively, relative to those of control hydrogel without MPC monomer. The results presented herein demonstrate the potential of expending biocompatible monomers such as HEMA and MPC to efficiently generate a biomaterial that possesses both bio‐membrane mimicking character and protein and bacterial resistant properties for various industrial and biomedical applications.

Bilayer Adsorption of Lysozyme on Nanodiamonds in Aqueous Suspensions

The interactions of one of the most famous enzymes, lysozyme, with carboxylated nanodiamonds in water were studied. It was found that stable complexes are formed as a result of lysozyme adsorption ...

Protein adsorption behaviors on siloxane-containing vaterite particles

The adsorption properties of two types of proteins, i.e., acidic bovine serum albumin (BSA) and basic lysozyme (LSZ), on siloxane-containing vaterite particles (SiV) were examined. SiV particles showing a positive zeta potential of 44 mV electrostatically adsorbed ~ 40 mg/g of negatively charged BSA in Tris buffer solution after soaking for 2 weeks. LSZ adsorption on SiV particles was 14 mg/g in 2 weeks, despite it being a positively charged protein. The adsorption of LSZ can be explained via the rapid dissolution of amorphous calcium carbonate phase and reprecipitation of positively charged calcite phase. SiV might be effective for designing biomaterials owing to high adsorption affinity for both, acidic as well as basic proteins.

A New Strategy for Reporting Specific Protein Binding Events at Aqueous-Liquid Crystal Interfaces in the Presence of Non-Specific Proteins.

Aqueous-liquid crystal (LC) interfaces offer promise as responsive interfaces at which biomolecular recognition events can be amplified into macroscopic signals. However, the design of LC interfaces that distinguish between specific and non-specific protein interactions remains an unresolved challenge. Herein, we report the synthesis of amphiphilic monomers, dimers, and trimers conjugated to sulfonamide ligands via triazole rings, their assembly at aqueous-LC interfaces, and the orientational response of LCs to the interactions of carbonic anhydrase II (CAII) and serum albumin with the oligomer-decorated LC interfaces. Of six oligomers synthesized, only dimers without amide methylation were found to assemble at aqueous interfaces of nematic 4-cyano-4'-pentylbiphenyl (5CB) to induce perpendicular LC orientations. At dimer-decorated LC interfaces, we found that concentrations of CAII less than 4 μM did not measurably perturb the LC but prevented non-specific adsorption and penetration of serum albumin into the dimer-decorated interface that otherwise triggered bright, globular LC optical domains. These experiments and others (including competitive adsorption of CAII, BSA, and lysozyme) support our hypothesis that specific binding of CAII to the dimer prevents LC anchoring transitions triggered by non-specific adsorption of serum albumin. We illustrate the utility of the approach by reporting (i) the relative activity of two small-molecule inhibitors (6-ethoxy-2-benzothiazolesulfonamide and benzenesulfonamide) of CAII to sulfonamide and (ii) proteolytic digestion of a protein (CAII) by thermolysin. Overall, the results in this paper provide new insight into the interactions of proteins at aqueous-LC interfaces and fresh ideas for either blocking non-specific interactions of proteins at surfaces or reporting specific binding events at LC interfaces in the presence of non-specific proteins.

Molecular simulations of lysozyme adsorption on an electrically responsive mixed self-assembled monolayer

Lysozyme adsorption on electrically responsive carboxyl/hydroxyl (SC10COOH/SC3OH) self-assembled monolayer (SAM) was investigated by molecular simulation. Results show that applied positive electric fields help to increase the surface hydrophilicity by aligning the chains parallel to the surface normal and exposing their carboxyl groups, whereas applied negative electric fields increase the surface hydrophobicity by bending the SC10COOH chains to expose their alkyl moieties. Accordingly, lysozyme behavior on the SAM was affected. Protein-surface interactions are significantly weakened by a strong negative electric field because of the decreased electrostatic interactions. However, the applied positive electric fields do not strengthen the protein-surface interactions probably because the hydrogen bonds formed within the surface chains, the binding of water molecules to form a water layer and the competitive adsorption from counterions can all decrease the interactions of the SAM with lysozyme. Adsorbed orientations of lysozyme could be regulated among “back-on”, “bottom-leaning” and “side-on” by applying a strong positive electric field or negative electric fields. Analysis of the conformational changes of lysozyme reveals that there is no direct relation between the protein deformation and the strength or direction of electric fields. This work may help to design and fabricate efficient electrically responsive surface in experiments for biotechnological applications.

Lipopolysaccharide increases lysozyme adhesion to chitin

Background. To study the effect of lipopolysaccharide on the adsorption of lysozyme on chitin.Methods. Egg white lysozyme, lipopolysaccharide and chitin were used. The lyophilized cells of M. lyzodeikticus were used as a lysozyme substrate. Lysozyme activity was determined by the bacteriolytic method using different concentrations (8-512 μg/ml). Lysozyme adsorption on chitin was determined by the loss of lysozyme from solution after shaking with chitin. The effect of lipopolysaccharide (0.6-40 μg/ml) on lysozyme adsorption was evaluated by the loss of lysozyme from the solution after shaking with 50 mg of chitin.Results. In determining the activity of lysozyme by the bacteriolytic method, the linear nature of the dependence of activity on the concentration of the enzyme is observed only at low concentrations (8-100 μg / ml). Lipopolysaccharide does not affect the bacteriolytic activity of lysozyme, however, dose-dependently increases the adsorption of lysozyme on chitin.Conclusion. Lipopolysaccharide increases the adsorption of lysozyme on chitin.

Effect of particle functionalization and solution properties on the adsorption of bovine serum albumin and lysozyme onto silica nanoparticles

Silica nanoparticles present an enormous potential as controlled drug delivery systems with high selectivity towards diseased cells. This application is directly related to the phenomenon of protein corona, characterized by the spontaneous adsorption of proteins on the nanoparticle surface, which is not fully understood. Here, we report an investigation on the influence of pH, ionic strength and temperature on the thermodynamics of interaction of bovine serum albumin protein (BSA) with non-functionalized silica nanoparticles (SiO2NPs). Complementary, we also investigated the ability of polyethylene glycol (PEG) and zwitterionic sulfobetaine (SBS) surface-modified nanoparticles to prevent the adsorption of BSA (protein negatively charged at physiological pH) and lysozyme (protein positively charged at physiological pH). We showed that BSA interaction with SiO2NPs is enthalpically governed. On the other hand, functionalization of silica nanoparticles with PEG and SBS completely prevented BSA adsorption. However, these functionalized nanoparticles presented a negative zeta potential and were not able to suppress lysozyme anchoring due to strong nanoparticle-protein electrostatic attraction. Due to the similarity of BSA with Human Serum Albumin, this investigation bears a resemblance to processes involved in the phenomenon of protein corona in human blood, producing information that is relevant for the future biomedical use of functionalized nanoparticles.

Insight into the binding and conformational changes of hemoglobin/lysozyme with bimetallic alloy nanoparticles using various spectroscopic approaches

This study explores the adsorption and binding interaction of bovine hemoglobin (BHb) and lysozyme (Lys) with gold/silver alloy nanoparticles (Au/Ag alloy NPs) using various spectroscopic approaches. The fluorescence behaviour of tryptophan residues of the proteins after Au/Ag alloy NPs addition revealed that the static quenching interactions. The changes in the morphology of Au/Ag alloy NPs upon BHb/Lys addition are characterized by HR-TEM analysis. Further, the conjugation of BHb/Lys with Au/Ag alloy NPs is evident from zeta potential and dynamic light scattering (DLS) techniques. The absorption profiles of Au/Ag alloy NPs with BHb/Lys resulted in the shift to the longer wavelength of surface plasmon resonance (SPR) band indicating the changes of refractive index around the NPs surface due to the proteins adsorption. The alteration in the structure of proteins with Au/Ag alloy NPs is evaluated using Fourier-transform infrared (FT-IR), absorption and circular dichroism (CD) measurements. The outcome of this study could be useful to understand the binding interaction of BHb/Lys proteins on Au/Ag alloy NPs and also to identify the health hazards and potential risks associated with Au/Ag alloy NPs.

Review of large-pore mesostructured cellular foam (MCF) silica and its applications

AbstractThe unique properties of mesostructured cellular foam (MCF) silica such as, large pore size, continuous three-dimensional (3D) pore system and hydrothermal robust materialallow favorable conditions for incorporating active sites to produce modified MCF silica as catalysts, biocatalysts and adsorbents. Recently, the modified MCF silicas were reported to be efficient catalysts for the hydrogenation of phenylacetylene, heck coupling reaction of arylboronic acid, etc. Biocatalysts derived from modified MCF silicas were found to be a potential to convert glucose to gluconic acid, hydrolysis of N-benzoyl-DLarginine-p-nitroanilide (BAPNA) and casein, transesterification of racemic 1 phenyl- ethanol and hydrolytic, etc. Several separation processes such as CO2 capture and adsorption of L-tryptophan, lysozyme and bovine serum were sucessfully conducted using adsorbents derived from modified MCF silicas. This paper reviews the synthesis of the MCF silica material and the incorporation of active sites or immobilization of enzymes in the MCF silica material. Additionally, a detailed understanding of the characterization of the modified MCF silicas, which includes pore size, active sites/enzymes sizes, amount of active sites/enzymes bound with the MCF silica, was also discussed to obtain their potentialities as catalysts, biocatalysts and adsorbents. The review also describes recent progress on the applications of the MCF silica.

PH-responsive polymer assisted aptamer functionalized magnetic nanoparticles for specific recognition and adsorption of proteins

A new adsorbent based on pH-responsive polymer assisted aptamer functionalized magnetic nanoparticles was developed for specific recognition and efficient adsorption of proteins. Arising from the synergistic effect of specific affinity of apatamer on protein and tunable hydrophobic/hydrophilic property of pH-responsive polymer, the adsorbent exhibited excellent adsorption capacity for target protein. Notably, because of the pH-responsive property of the polymer, the adsorption and desorption process could be regulated through varying environmental pH. The resultant adsorbent that immobilized with lysozyme binding aptamer was successfully applied in specific recognition and efficient adsorption of lysozyme in egg white samples and good recovery results in the range of 95.2–103.2% were obtained. Moreover, the adsorbent immobilized with cytochrome C binding aptamer also exhibited satisfactory adsorption to cytochrome C. The synergistic effect of pH-responsive polymer and aptamer promoted the recognition selectivity and adsorption capacity to target protein, illustrating a facile way for construction of more specific protein adsorbents.

Calorimetric approach to understand pH and salt influence on the adsorption mechanism of lysozyme to a traditional cation exchanger

Built upon our interest in illustrating the complexity of protein adsorption onto chromatographic supports and to understand the rule of nonspecific interactions in the ion exchange adsorption process, a traditional model system (lysozyme – carboxymethyl cellulose) was used to determine the charge influence during biomolecule adsorption. Flow microcalorimetry (FMC) was exploit as a dynamic technique that provides adsorption and desorption heat signals for a specific system, permitting an improved understanding of the driving forces and mechanisms involved during the interaction. For this purpose, measurements were made at pH 8 at both absence and presence of salt (NaCl 50 mM) and compared with previous studies performed at pH 5. Distinct FMC profiles were observed regarding pH. For most of the experiments, two exothermic heat signals are observed at pH 8 while at pH 5 one endothermic and one exothermic peak are shown. This difference was justified with a less energy demanding for desolvation at pH 8. Lysozyme adsorption was shown to be a multi-step process involving desolvation, primary protein adsorption and secondary adsorption after reorientation with distinct contributions to the overall energy. At pH 8, the exothermic contribution to the adsorption process is lower compared to pH 5, which is justified by the lower charge density that lysozyme presents at pH 8 compared to pH 5.

Assessment of a new dual effective combo polymer structure for separation of lysozyme from hen egg white

Water-soluble O-carboxymethyl chitosan (O-CMCS) Schiff base granules were prepared, and Cu2+ ions were decorated. Then, dual effective combo cryogel structures were prepared by free radical polymerization using Cu2+-O-CMCS granules as embedding agent. 2-Hydroxyethyl methacrylate and N,N′-methylene-bis-acrylamide were used as monomer and cross-linker pair, respectively. Prepared combo structures were characterized by FTIR, SEM and swelling tests measurements, and investigated for lysozyme adsorption. Maximum lysozyme adsorption was observed in phosphate buffer of pH 8.0 as 103.3 mg/g with initial lysozyme concentration of 3 mg/mL. On the other hand, the most efficient adsorption was carried out at 0.5 mL/min, at 25 °C. It was also observed that combo cryogels could be repeatedly used for adsorption and desorption cycles of lysozyme molecules. The purification efficiency of adsorbent was highlighted as 83.4% with 88.2% purification yield from hen egg white via sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE).

Immobilization of surface non-affinitive protein onto a metal surface by an external electric field

We investigated an alternate technique to coat the surface with a protein having no surface affinity, without the use of any exotic chemical agents. An external electric field was utilized to prepare the protein coating on a metal substrate. Stainless steel (St) substrate and lysozyme (LSZ) were used as the surface to be coated and the model non-adsorptive protein, respectively. Dynamics of the adsorption of LSZ on the St surface in the presence and absence of an external electric potential (EEP) were monitored by in-situ ellipsometry. Applying negative surface potential (-0.4V vs Ag/AgCl) forced the adsorption of LSZ onto the St surface where LSZ did not adsorb without applying any EEP. The repetition of the EEP-application and -cut-off indicated the controllability of the LSZ coating amount depending on the total duration of the EEP-application. The coated LSZ largely remained bound to the surface even by the cut-off of the external electric field, the ratio of which to the detached amount was roughly constant (approximately 7:3). Furthermore, the LSZ coated surface on the St substrate was found to be reversibly switched between being affinitive and non-affinitive to a typical model protein adsorbate (bovine serum albumin) by the EEP-application and cut-off.

Protein cation exchangers derived by charge reversal from poly(ethylenimine)-Sepharose FF: Comparisons between two derivatization routes

It has been known that anion exchangers prepared by grafting poly(ethyleneimine) (PEI) onto Sepharose FF (PEI-Sepharose) at ionic capacities (IC) over 600 mmol/L show both high protein adsorption capacity and uptake kinetics, and charge reversal of PEI-Sepharose by modification with succinic anhydride can produce protein cation exchangers of high capacity and uptake rate. Previously, a Charge Reversal-then-Reduction procedure (route A) was studied for preparation of cation exchangers of different IC values from PEI-Sepharose. In this work, we proposed a new route, that is, Charge Reduction-then-Reversal route (route B), to develop cation exchangers of different IC values from PEI-Sepharose FF with an IC of 700 mmol/L (FF-PEI-L700) as the starting resin. The two kinds of cation exchangers (route A, PEI-L700-CRn; route B, PEI-Rm-Cn) are compared for lysozyme (Lys) adsorption and chromatography. The two modification routes result in the difference in the ligand structures that significantly affect protein adsorption equilibrium and kinetics. Route A introduces long electroneutral groups that hinder protein adsorption and reduce equilibrium capacity. Moreover, charge reversal by reaction with succinic anhydride could cause diamide formation, which reduces remaining carboxyl groups or the IC. In the charge-reduced FF-PEI-Rm resins of the lowest IC (394 mmol/L) prepared in route B, the diamide formation was little due to the lack of primary and secondary amine groups, so its charge reversal makes a higher-IC cation exchanger. This makes PEI-Rm-Cn show a higher IC (589 mmol/L) than PEI-L700-CRn (463 mmol/L) in which De/D0 jumps about four times. The differences in the adsorption equilibrium and kinetics make the two kinds of resins behave distinctly in dynamic adsorption and chromatography. Namely, PEI-Rm-Cn resins display obviously higher dynamic binding capacities than PEI-L700-CRn resins in the IC range studied. For instance, the DBC (at 10% breakthrough) of PEI-R590-C680 (192 mg/mL) is 33% higher than that of PEI-L700-CR680 (144 mg/mL). This is proved by the purification of Lys from chicken egg white solution, in which the PEI-R590-C680 column purified Lys with a recovery yield 14% higher than the PEI-L700-CR680 column. This research thus demonstrated that Charge Reduction-then-Reversal route is superior over Charge Reversal-then-Reduction route in fabricating a high-capacity cation exchanger from PEI-Sepharose.

Prediction of the elution profiles of proteins in mixed salt systems in hydrophobic interaction chromatography

Hydrophobic interaction chromatography (HIC) is often used for purifying proteins. A mathematical model to describe the complex effects of salts on the adsorption equilibria in HIC has recently been introduced by our group. It describes not only the influence of single salts, but also salt mixtures, in which cooperative effects may occur. The influence of the salts is thereby modeled with a Taylor series expansion in the individual ion molarities. In the present study, the model of the adsorption equilibrium is coupled with a lumped kinetic model of the adsorption kinetics to obtain a model of the elution of proteins in HIC adsorption columns. The column model is tested using experimental data on the adsorption of bovine serum albumin (BSA) and lysozyme (LYS) on the mildly hydrophobic resin Toyopearl PPG-600M at pH 7. The studied salts are ammonium chloride, sodium chloride, ammonium sulfate, and sodium sulfate as well as binary and ternary mixtures of them. The parameters of the lumped kinetic model are protein-specific and were fitted to the elution profiles of the single proteins in presence of single salts. The model was then used to predict the elution profiles of BSA and LYS solutions containing both proteins, for single salts as well as for binary and ternary salt mixtures. Both isocratic and gradient elution were studied. Furthermore, the model was applied to identify the optimal overall ionic strength for the separation of the two proteins by isocratic elution and the optimal linear gradient of the salt concentration in a multicriteria approach where the conflicting goals are high separation yield and low elution volume.

Competitive adsorption of bovine serum albumin and lysozyme on a beta-tricalcium phosphate nanocoating

The competitive and cooperative adsorptions of proteins on the surface of implanted materials can affect the biocompatibility and osteogenesis of the material. The adsorption processes are related to the properties of the materials, as well as to the nature of the proteins. Competitive adsorption of bovine serum albumin (BSA) and lysozyme (LSZ), both sequentially, and from a mixture, was investigated on a beta-tricalcium phosphate (β-TCP) nanocoating using quartz crystal microbalance with dissipation technique. In experiments with individual proteins, the BSA adlayer showed higher adsorption quality and viscoelasticity than the LSZ on the β-TCP surface. In the sequential adsorption of LSZ + BSA, protein exchange was observed but not in the BSA + LSZ sequence, because of the larger molecular and higher surface affinity of BSA relative to LSZ. The substitution rate increased as the increasing concentration of the BSA solution. With a BSA&LSZ mixture, the total amount of protein adsorbed was greater than that for individual or sequential adsorption, owing to the formation of BSA and LSZ agglomerates. The electrostatic force and affinity were dominant factors governing the adsorption behavior between BSZ and LSZ on the β-TCP surface. This work will be beneficial to understand the protein adsorption mechanism on β-TCP surface and provide information to design better osteogenic materials.