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

Marta Ziegler-Borowska,Pawel Nowak,Kinga Mylkie,Adam Sikora,Anna Kaczmarek-Kedziera,Dorota Chelminiak-Dudkiewicz,Patryk Rybczynski

doi:10.3390/molecules25081945

Marta Ziegler-Borowska, Pawel Nowak + Show 5 more

Open Access

https://doi.org/10.3390/molecules25081945

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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.

Highlights

Human serum albumin (HSA) is one of the most important globular plasma proteins containing a single heart-shaped polypeptide chain of 585 amino acid residues organized in three homologous domains (I, II and III) with two subdomains, A and B (Figure 1)
Drugs related to HSA are pharmacologically inactive—only the free, unbound fraction of the drug is active and reaching the target causes a pharmacological response of the body and is available to elimination processes [3]
Because the plasma HSA content is high, the study of the interaction of drugs with this protein is very important in pharmacology, especially for newly tested active substances with expected pharmacological activity

Summary

Introduction

Human serum albumin (HSA) is one of the most important globular plasma proteins containing a single heart-shaped polypeptide chain of 585 amino acid residues organized in three homologous domains (I, II and III) with two subdomains, A and B (Figure 1). The most important role of HSA is the ability to bind and transport substances [1]. Because the plasma HSA content is high, the study of the interaction of drugs with this protein is very important in pharmacology, especially for newly tested active substances with expected pharmacological activity. There are so many models describing the interaction of HSA with a drug that for each active substance able to bind to albumin this interaction must be qualitatively and quantitatively determined

Objectives

Methods

Results