%0 Journal Article
%T Immobilization of Homogeneous Monomeric, Oligomeric and Fibrillar A¦Â Species for Reliable SPR Measurements
%A Daniel Frenzel
%A Julian M. Gl¨¹ck
%A Oleksandr Brener
%A Filipp Oesterhelt
%A Luitgard Nagel-Steger
%A Dieter Willbold
%J PLOS ONE
%D 2014
%I Public Library of Science (PLoS)
%R 10.1371/journal.pone.0089490
%X There is strong evidence that the amyloid-beta peptide (A¦Â) plays a central role in the pathogenesis of Alzheimer's disease (AD). In this context, a detailed quantitative description of the interactions with different A¦Â species is essential for characterization of physiological and artificial ligands. However, the high aggregation propensity of A¦Â in concert with its susceptibility to structural changes due to even slight changes in solution conditions has impeded surface plasmon resonance (SPR) studies with homogeneous A¦Â conformer species. Here, we have adapted the experimental procedures to state-of-the-art techniques and established novel approaches to reliably overcome the aforementioned challenges. We show that the application of density gradient centrifugation (DGC) for sample purification and the use of a single chain variable fragment (scFv) of a monoclonal antibody directed against the amino-terminus of A¦Â allows reliable SPR measurements and quality control of the immobilized A¦Â aggregate species at any step throughout the experiment.
%U http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0089490