Preparation of protein nanoparticle by dynamic aggregation and ionizing-induced crosslinking

Albumin-based nanoparticle trehalose lyophilisation stress-down to preserve structure/function and enhanced binding

“…We think the binding pocket where Tyr is present may be not available to interact with MC540 in accordance with other authors [15]. If this were not to be true, then differences between data obtained when the excitation is at 295 nm and/or at 280 nm from the quenching curves should have been observed.…”

“…Table 4 depicts the maxima drug loaded for the BSA NP lyophilised and its dimer. No difference were observed, both showed the same B mx value (5.31E − 6 ± 0.28E − 6 The drug loaded by the lyophilised NP is significantly less than the one loaded by the BSA NP non lyophilised; 0.9E − 4 ± 0.3E − 4 [15]. This could be due to the trehalose present in solution, preventing drug loading.…”

“…When comparing BSA NP/MC540 interaction Kd value (2.76E − 4 ± 1.34E − 4), from Achilli et al [15] with that of the, BSA NP lyophilised with trehalose, the K d value obtained in this study has a greater affinity (hundred-fold) for the probe. Reasons may be that after lyophilisation the NP acquires a new depth on hydrophobicity.…”

“…The fitting performed on the curve (Fig. 5b) describes the binding process to be a one-step process similar to the one between MC540 and BSA NP non lyophilised [15]. Therefore, the binding constants K d and B mx (Table 4) from Scatchard plot (Fig.…”

“…35 nm from TEM measurements and previous findings [2,15]. Besides, based on microscopies (SEM and TEM) and on Ericksonís paper [14], an approximate calculation of the molecular weight of the NP can be calculated using equations assuming that the NP has a spherical shape the volume is: V = 4/3R 3 .…”