Optimization of layering technique and secondary structure analysis during the formulation of nanoparticles containing lysozyme by quality by design approach

Abstract

In our study, core-shell nanoparticles containing lysozyme were formulated with precipitation and layering self-assembly. Factorial design (DoE) was applied by setting the process parameters during the preparation with Quality by Design (QbD) approach. The factors were the concentration of lysozyme and sodium alginate, and pH. Our aim was to understand the effect of process parameters through the determination of mathematical equations, based on which the optimization parameters can be predicted under different process parameters. The optimization parameters were encapsulation efficiency, particle size, enzyme activity and the amount of α-helix structure. The nanoparticles were analysed with transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR) and circular dichroism (CD) spectroscopy. Based on our results, we found that pH was the most important factor and pH 10 was recommended during the formulation. Enzyme activity and α-helix content correlated with each other very well, and particle size and encapsulation efficiency also showed very good correlation with each other. The results of the α-helix content of FTIR and CD measurements were very similar for the precipitated lysozyme due to the solid state of lysozyme. The mixing time had the best influence on the encapsulation efficiency and the particle size, which leads to the conclusion that a mixing time of 1 h is recommended. The novelty in our study is the presentation of a mathematical model with which the secondary structure of the protein and other optimization parameters can be controlled in the future during development of nanoparticle based on the process parameters.

Quality by Design-Based Development of Solid Self-Emulsifying Drug Delivery System (SEDDS) as a Potential Carrier for Oral Delivery of Lysozyme

“…A high positive coefficient was also observed in the case of pH value (Coeff. = +7.46), indicating higher enzyme activity at higher pH values, which is in line with previously published results [ 47 ]. The two-way interaction coefficient (x 1 x 2 ) was also high (Coeff.…”

Preparation and optimization of hydrophobic ion pairing complex of lysozyme

“…Hydrophobic ion pairing (HIP) has emerged as an approach to enhance the encapsulation and loading of water soluble peptides/proteins in their micro and nanocarriers. The hydrophobicity of these molecules is increased through a reversible electrostatic complexation with an oppositely charged amphipathic molecule at suitable pH [1,2]. This work aims to investigate different factors that affect the preparation of HIP of lysozyme (LYZ) with the surfactant sodium dodecyl sulphate (SDS).…”