It is known that there are dimmers and hexamers of BSA on aqueous solutions and this quaternary structure is probably involved during the adsorption process at high concentration [4]. For all phosphate or acetate buffers concentrations used in this work, the adsorption process did not Lapatinib molecular weight follow a typical Langmuir equation as observed elsewhere [19], probably because the adsorption process involved protein-protein interactions. For 0.01 M buffer concentration – phosphate or acetate – a good agreement to the data points was obtained by fitting the adsorption isotherms with a Langmuir–Freundlich equation as shown in Fig. 2a and b and Table

1. The nature of buffer did not influence the adsorption efficiency (am and K parameters). The positive values of r higher than unity indicated the existence of a positive cooperativity in the protein–protein interaction. The increase of the buffer concentration induced an enhancement of the cooperative interaction between BSA molecules and a decrease of the maximum adsorption sites. This behavior could be explained by the competition between phosphate Romidepsin in vitro groups (buffer) and COOH groups from BSA for HA surface sites as showed by

Wessel et al. [19] and Yin et al. [18]. The maximum binding sites and the affinity constant (am = 1.5 mg/m2 and K = 2.5 mL/mg, considering HA BET area of 45 m2/g) of the HA used in this work were higher than those obtained by Wassel et al. [19] (am = 0.62 mg/m2 and K = 0.15 mL/mg) using a calcium deficient HA with specific surface

area of 26 m2/g and a Langmuir fitting. The higher values of am and K parameters verified in this work may be explained by the composition and stoichiometry of the HA surface. The BSA adsorption on HA showed a complex kinetics pattern when phosphate buffer increased to 0.05 M. In this condition the data points could not be fitted by a single equation for all protein initial concentrations. As shown in Fig. 2c the best simulation was achieved by using a Langmuir–Freundlich equation for BSA concentration below Non-specific serine/threonine protein kinase 0.4 mg/mL and a pure Freundlich equation for BSA concentration above 0.4 mg/mL. Some conclusions could be taken from these results: (i) the increase of buffer concentration leads to a decrease in adsorption and binding affinity, (ii) a typical Langmuir behavior, associated to a protein monolayer formation and a HA surface with homogeneous distributed sites was not found, (iii) the occurrence of a Langmuir–Freundlich mechanism suggested the existence of cooperative protein-protein interaction on HA surface even for low concentration of BSA, (iv) at high protein concentration the interaction between BSA molecules predominated at HA surface. Many aspects might be considered to understand the non-typical Langmuir adsorption process obtained in this work.