Stable, ligand-doped, poly(bis-SorbPC) lipid bilayer arrays for protein binding and detection

James R. Joubert, Kathryn A. Smith, Erin Johnson, John P. Keogh, Vicki H. Wysocki, Bruce K. Gale, John C. Conboy, S. Scott Saavedra

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

A continuous-flow microspotter was used to generate planar arrays of stabilized bilayers composed of the polymerizable lipid bis-SorbPC and dopant lipids bearing ligands for proteins. Fluorescence microscopy was used to determine the uniformity of the bilayers and to detect protein binding. After UV-initiated polymerization, poly(lipid) bilayer microarrays were air-stable. Cholera toxin subunit b (CTb) bound to an array of poly(lipid) bilayers doped with GM1, and the extent of binding was correlated to the mole percentage of GM1 in each spot. A poly(lipid) bilayer array composed of spots doped with GM1 and spots doped with biotin-DOPE specifically bound CTb and streptavidin to the respective spots from a dissolved mixture of the two proteins. Poly(bis-SorbPC)/GM1 arrays retained specific CTb binding capacity after multiple regenerations with a protein denaturing solution and also after exposure to air. In addition, these arrays are stable in vacuum, which allows the use of MALDI-TOF mass spectrometry to detect specifically bound CTb. This work demonstrates the considerable potential of poly(lipid) bilayer arrays for high-throughput binding assays and lipidomics studies.

Original languageEnglish (US)
Pages (from-to)1310-1315
Number of pages6
JournalACS Applied Materials and Interfaces
Volume1
Issue number6
DOIs
StatePublished - Jun 24 2009

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Keywords

  • GM1
  • MALDI-TOF MS
  • cholera toxin
  • continuous-flow microspotter
  • microarray
  • poly(lipid)
  • supported lipid bilayer

ASJC Scopus subject areas

  • Materials Science(all)

Cite this

Joubert, J. R., Smith, K. A., Johnson, E., Keogh, J. P., Wysocki, V. H., Gale, B. K., Conboy, J. C., & Saavedra, S. S. (2009). Stable, ligand-doped, poly(bis-SorbPC) lipid bilayer arrays for protein binding and detection. ACS Applied Materials and Interfaces, 1(6), 1310-1315. https://doi.org/10.1021/am900177p