4.3 Article

Physical elution in phage display selection of inorganic-binding peptides

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.msec.2008.05.003

Keywords

Phage display; Physical elution; Ultrasonication; Inorganic-binding peptides; Muscovite mica

Funding

  1. Turkish State Planning Organization
  2. US Army Research Office-Defense University Research initiative in Nanotechnology [DAAD19-01-1-04999]
  3. Genetically Engineered Materials Science and Engineering Center
  4. NSF-MRSEC at the University of Washington

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Phage display is a commonly utilized in vivo approach in selecting peptides specific to solid inorganic materials. In this process, traditionally, high affinity peptides are recovered by a chemical elution method, which involves contacting the phage library with the desired inorganic, washing the weak binders. and eluting the tight binders under harsh buffer conditions. This process may result in incomplete removal of all strong binders, separation of the phage from the display protein, or may modify the material surface. To overcome these potential limitations, we developed a physical elution technique based on ultrasonication. Here, we report two optimized ultrasonication protocols by which we selected peptides specific to natural mineral mica. We first performed a 30-s physical elution after the chemical elution step and increased the efficiency of screening strong binders by about 100%. Encouraged by the results, we applied physical elution-only protocol where we obtained 45% of the selected sequences as strong binders. The approach has a far shorter total elution time, i.e., seconds compared to hours in traditional chemical elution. The novel physical elution approach using ultrasonication reported herein can be a highly efficient alternate step in the screening of solid material specific peptides. (C) 2008 Elsevier B.V. All rights reserved.

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