Measuring binding of protein to gel-bound ligands using magnetic levitation.

Journal of the American Chemical Society
Nathan D ShapiroGeorge M Whitesides

Abstract

This paper describes the use of magnetic levitation (MagLev) to measure the association of proteins and ligands. The method starts with diamagnetic gel beads that are functionalized covalently with small molecules (putative ligands). Binding of protein to the ligands within the bead causes a change in the density of the bead. When these beads are suspended in a paramagnetic aqueous buffer and placed between the poles of two NbFeB magnets with like poles facing, the changes in the density of the bead on binding of protein result in changes in the levitation height of the bead that can be used to quantify the amount of protein bound. This paper uses a reaction-diffusion model to examine the physical principles that determine the values of rate and equilibrium constants measured by this system, using the well-defined model system of carbonic anhydrase and aryl sulfonamides. By tuning the experimental protocol, the method is capable of quantifying either the concentration of protein in a solution, or the binding affinities of a protein to several resin-bound small molecules simultaneously. Since this method requires no electricity and only a single piece of inexpensive equipment, it may find use in situations where portability and ...Continue Reading

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Citations

Jun 13, 2012·Analytical Chemistry·Nathan D ShapiroGeorge M Whitesides
Jan 28, 2016·Analytical Chemistry·Alex NemiroskiGeorge M Whitesides
Jan 2, 2016·Journal of the American Chemical Society·Alex NemiroskiGeorge M Whitesides
Aug 27, 2015·PloS One·Stephanie KnowltonSavas Tasoglu
Dec 5, 2014·Lab on a Chip·Anand Bala SubramaniamGeorge M Whitesides
Apr 28, 2018·Biomaterials Science·Esra TürkerAhu Arslan-Yildiz
Dec 17, 2014·Angewandte Chemie·Antara PalAndrei V Petukhov
Apr 13, 2013·Chemical Society Reviews·Christopher J GraySabine L Flitsch
Jan 22, 2020·Nanoscale·Ali Akbar AshkarranMorteza Mahmoudi
Jun 6, 2019·Angewandte Chemie·Shencheng GeGeorge M Whitesides
Jun 11, 2015·Advanced Materials·Savas TasogluUtkan Demirci
Jan 16, 2015·Advanced Materials·Jonathan W HennekGeorge M Whitesides
Sep 19, 2017·Nanoscale·Vikash MalikPeter Schurtenberger
Apr 15, 2015·Advanced Healthcare Materials·Savas TasogluUtkan Demirci
Aug 14, 2013·Angewandte Chemie·Manza B J AtkinsonGeorge M Whitesides
Jul 1, 2017·Advanced Functional Materials·Wujun ZhaoLeidong Mao
Aug 27, 2013·Analytical Chemistry·David K BwambokGeorge M Whitesides
Dec 7, 2017·Journal of the American Chemical Society·Shencheng GeGeorge M Whitesides
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Jun 12, 2018·Journal of the American Chemical Society·Shencheng GeGeorge M Whitesides
Nov 20, 2021·Analytical Chemistry·Haonan LiQiang Liao
Oct 15, 2021·Angewandte Chemie·Parthena Chorti, Dionysios C Christodouleas

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