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Partial-filling affinity capillary electrophoresis and quartz crystal microbalance with adsorption energy distribution calculations in the study of biomolecular interactions with apolipoprotein E as interaction partner

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Abstract

Adsorption energy distribution (AED) calculations were successfully applied to partial-filling affinity capillary electrophoresis (PF-ACE) to facilitate more detailed studies of biomolecular interactions. PF-ACE with AED calculations was employed to study the interactions between two isoforms of apolipoprotein E (apoE) and dermatan sulfate (DS), and a quartz crystal microbalance (QCM) was used in combination with AED calculations to examine the interactions of the 15-amino-acid peptide fragment of apoE with DS. The heterogeneity of the interactions was elucidated. Microscale thermophoresis was used to validate the results. The interactions studied are of interest because, in vivo, apolipoprotein E localizes on DS-containing regions in the extracellular matrix of human vascular subendothelium. Two-site binding was demonstrated for the isoform apoE3 and DS, but only one-site binding for apoE2–DS. Comparable affinity constants were obtained for the apoE2–DS, apoE3–D3, and 15-amino-acid peptide of apoE–DS using the three techniques. The results show that combining AED calculations with modern biosensing techniques can open up another dimension in studies on the heterogeneity and affinity constants of biological molecules.

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Acknowledgments

Emilia Danilowich-Luebert is thanked for her help with the MST measurements. Financial support was provided by the Research Council for Natural Sciences and Engineering, the Academy of Finland (under grants 1133184 (K. L., G.C.-K and M.-L.R.) and 267497 / 273645 (M. K.)), the Academy of Finland under grant 257545 (M. J.), and the Swedish Research Council (VR) under grant 621-2012-3978 (J.S. and T.F.).

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Authors and Affiliations

  1. Laboratory of Analytical Chemistry, Department of Chemistry, University of Helsinki, P.O. Box 55, 00014, Helsinki, Finland

    Katriina Lipponen, Sari Tähkä, Geraldine Cilpa-Karhu & Marja-Liisa Riekkola

  2. Department of Engineering and Chemical Sciences, Karlstad University, 651 88, Karlstad, Sweden

    Jörgen Samuelsson & Torgny Fornstedt

  3. National Institute for Health and Welfare and FIMM, Institute of Molecular Medicine Finland, Biomedicum, Haartmaninkatu 8, 00290, Helsinki, Finland

    Matti Jauhiainen & Jari Metso

  4. Biohybrid Materials, Department of Biotechnology and Chemical Technology, Aalto University, 00076, Espoo, Finland

    Mauri Kostiainen

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  1. Katriina Lipponen
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Correspondence to Marja-Liisa Riekkola.

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Lipponen, K., Tähkä, S., Samuelsson, J. et al. Partial-filling affinity capillary electrophoresis and quartz crystal microbalance with adsorption energy distribution calculations in the study of biomolecular interactions with apolipoprotein E as interaction partner. Anal Bioanal Chem 406, 4137–4146 (2014). https://doi.org/10.1007/s00216-014-7821-9

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