Characterization of binding by sulfonylureas with normal or modified human serum albumin using affinity microcolumns prepared by entrapment.

Saumen Poddar, Ashley G Woolfork, Sazia Iftekhar, Susan T Ovbude, David S Hage

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 2023 Jul 15

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Modification of proteins can occur during diabetes due to the formation of advanced glycation end-products (AGEs) with reactive dicarbonyls such as glyoxal (Go) and methylglyoxal (MGo). Human serum albumin (HSA) is a serum protein that binds to many drugs in blood and that is known to be modified by Go and MGo. This study examined the binding of various sulfonylurea drugs with these modified forms of HSA by using high-performance affinity microcolumns prepared by non-covalent protein entrapment. Zonal elution experiments were employed to compare the retention and overall binding constants for the drugs with Go- or MGo-modified HSA vs normal HSA. The results were compared to values from the literature, such as measured or estimated using affinity columns containing covalently immobilized HSA or biospecifically-adsorbed HSA. The entrapment-based approach provided estimates of global affinity constants within 3-5 min for most of the tested drugs and with typical precisions of ±10-23%. Each entrapped protein microcolumn was stable for over at least 60-70 injections and one month of use. The results obtained with normal HSA agreed at the 95% confidence level with global affinity constants that have been reported for the given drugs in the literature. It was found for HSA that had been modified with clinically-relevant levels of either Go or MGo that an increase in the global affinity constant of up to 2.1-fold occurred for some of the tested drugs. The information acquired in this study can be used in the future to adapt this entrapment-based approach to study and evaluate interactions between other types of drugs and normal or modified binding agents for clinical testing and biomedical research. Copyright © 2023 Elsevier B.V. All rights reserved.

Citation

Saumen Poddar, Ashley G Woolfork, Sazia Iftekhar, Susan T Ovbude, David S Hage. Characterization of binding by sulfonylureas with normal or modified human serum albumin using affinity microcolumns prepared by entrapment. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 2023 Jul 15;1226:123798