Step by Step Preparation and Optimization of Lysozyme Hydrophobic Ion Pairing Complex

Abstract

Clinical applications of therapeutically attractive peptides/proteins have been delayed due to several challenges including chemical and physiological barriers. Different approaches were employed to overcome these barriers, for example, by improving the encapsulation of proteins into nanocarriers by hydrophobic ion pairing (HIP) complexation. In this approach, ionizable functional groups of proteins are engaged in electrostatic interactions with a counterion containing one or more hydrophobic moieties resulting in an increased hydrophobicity of such hydrophilic macromolecules and therefore, improving their encapsulation into nanocarriers using, for example, emulsification techniques (Ristroph et al., 2021). A naturally occurring cationic single chain polypeptide lysozyme (LYZ), which is known for its antimicrobial activity, has been commonly employed as a model peptide in nonparenteral dosage forms. In different studies, HIP complex of LYZ with surfactants such as sodium dodecyl sulphate (SDS) was prepared. Variable complexation efficiencies were obtained in these reports as different levels of factors affecting the complexation process were used. For instance, variable pH values of the used mediums were proposed for the optimum complexation (Asuman Bozkır, 2015; Yoo et al., 2001). This study aimed to prepare and optimize the HIP complex of LYZ and SDS with the help of the quality by design (QbD) approach. Risk assessment (RA), as one of the QbD tools, was applied to identify and prioritize different parameters influencing the preparation of such complex. Мaterials and methods