Testing for Ketoprofen Binding to HSA Coated Magnetic Nanoparticles under Normal Conditions and after Oxidative Stress

Abstract

Binding and transport of ligands is one of the most important functions of human blood serum proteins. Human serum albumin is found in plasma at the highest concentration. Because of this, it is important to study protein–drug interactions for this albumin. Since there is no single model describing this interaction, it is necessary to measure it for each active substance. Drug binding should also be studied in conditions that simulate pathological conditions of the body, i.e., after oxidative stress. Due to this, it is expected that the methods for testing these interactions need to be easy and fast. In this study, albumin immobilized on magnetic nanoparticles was successfully applied in the study of protein–drug binding. Ketoprofen was selected as a model drug and interactions were tested under normal conditions and artificially induced oxidative stress. The quality of obtained results for immobilized protein was confirmed with those for free albumin and literature data. It was shown that the type of magnetic core coverage does not affect the quality of the obtained results. In summary, a new, fast, effective, and universal method for testing protein–drug interactions was proposed, which can be performed in most laboratories.

Polymer-Coated Magnetite Nanoparticles for Protein Immobilization

“…Chitosan-coated nanoparticles can easily penetrate cell membranes, which is often used in biomedical research. Free hydroxyl and amine groups of chitosan allow the surface modification of nanoparticles [ 182 , 183 , 184 , 185 ]. In an acidic environment, free CS amino groups gain a positive charge, thanks to which they can react with negatively charged groups of nucleic acids, which are used, among others, in MRI imaging [ 186 ].…”

“…Later, Ziegler-Borowska and co-authors prepared magnetite nanoparticles coated with chemically-modified chitosan rich in free amino groups remote from the polymer chain ( Figure 19 ) [ 183 , 185 , 198 , 199 , 200 , 201 , 202 ]. First, chitosan-coated magnetite nanoparticles were synthesized by an in situ co-precipitation reaction in an alkaline solution.…”

“…Then, the reactive chitosan groups, hydroxyl and amino, were used for its functionalization, obtaining three polymer coatings with different content of amino groups able to bind protein ( Figure 19 ). Prepared nanoparticles were used for the chemical immobilization of proteins: lipase from Candida rugosa [ 183 , 193 , 201 ], human serum albumin (HSA) [ 185 , 199 ], and the androgen receptor (AR) [ 202 ].…”

“…Finally, these materials were used for HSA immobilization. Moreover, the influence of the selected method of functionalization of the polymer coating covering magnetite nanoparticles (MNP) as well as drying methods of the carrier on the immobilization of human serum albumin HSA was performed [ 185 ]. Albumin was immobilized on three types of nanoparticles coated with aminated chitosan with different content of amino groups at a large distance from the Fe 3 O 4 -CS-Et(NH 2 ) 1–3 surface.…”

“…The amount of HSA immobilized on the nanoparticles was about 149.96 mg HSA/g of the carrier, and the protein retained its activity after immobilization. Next, these HSA-aminated starch-coated MNPs were used for HSA-ketoprofen binding study [ 185 ].…”