Show simple item record

dc.contributor.authorShapiro, Nathan D.
dc.contributor.authorMirica, Katherine A.
dc.contributor.authorSoh, Siowling
dc.contributor.authorPhillips, Scott T.
dc.contributor.authorTaran, Olga
dc.contributor.authorMace, Charles R.
dc.contributor.authorShevkoplyas, Sergey S.
dc.contributor.authorWhitesides, George M.
dc.date.accessioned2014-03-17T20:38:36Z
dc.date.issued2012
dc.identifier.citationShapiro, Nathan D., Katherine A. Mirica, Siowling Soh, Scott T. Phillips, Olga Taran, Charles R. Mace, Sergey S. Shevkoplyas, and George M. Whitesides. 2012. Measuring Binding of Protein to Gel-Bound Ligands Using Magnetic Levitation. Journal of the American Chemical Society 134, no. 12: 5637–5646.en_US
dc.identifier.issn0002-7863en_US
dc.identifier.issn1520-5126en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:11931826
dc.description.abstractThis 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 low cost are important, such as in bioanalysis in resource-limited settings, point-of-care diagnosis, veterinary medicine, and plant pathology. It still has several practical disadvantages. Most notably, the method requires relatively long assay times and cannot be applied to large proteins (>70 kDa), including antibodies. The design and synthesis of beads with improved characteristics (e.g., larger pore size) has the potential to resolve these problems.en_US
dc.description.sponsorshipChemistry and Chemical Biologyen_US
dc.language.isoen_USen_US
dc.publisherAmerican Chemical Societyen_US
dc.relation.isversionofdoi:10.1021/ja211788een_US
dash.licenseOAP
dc.titleMeasuring Binding of Protein to Gel-Bound Ligands Using Magnetic Levitationen_US
dc.typeJournal Articleen_US
dc.description.versionAccepted Manuscripten_US
dc.relation.journalJournal of the American Chemical Societyen_US
dash.depositing.authorWhitesides, George M.
dc.date.available2014-03-17T20:38:36Z
dc.identifier.doi10.1021/ja211788e*
workflow.legacycommentsFAR 2013en_US
dash.contributor.affiliatedWhitesides, George
dc.identifier.orcid0000-0001-9451-2442


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record