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, Steven S Saavedra

Research output: Contribution to journalArticle

18 Citations (Scopus)

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

Fingerprint

Lipid bilayers
Cholera Toxin
Ligands
Proteins
Lipids
Bearings (structural)
Streptavidin
Fluorescence microscopy
Microarrays
Biotin
Air
Mass spectrometry
Assays
Polymerization
Doping (additives)
Throughput
Vacuum
1,2-bis(10(2',4'-hexadienoyloxy)decanoyl)-sn-glycero-3-phosphocholine
Protein Binding

Keywords

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

ASJC Scopus subject areas

  • Materials Science(all)

Cite this

Stable, ligand-doped, poly(bis-SorbPC) lipid bilayer arrays for protein binding and detection. / Joubert, James R.; Smith, Kathryn A.; Johnson, Erin; Keogh, John P.; Wysocki, Vicki H.; Gale, Bruce K.; Conboy, John C.; Saavedra, Steven S.

In: ACS Applied Materials and Interfaces, Vol. 1, No. 6, 24.06.2009, p. 1310-1315.

Research output: Contribution to journalArticle

Joubert, JR, Smith, KA, Johnson, E, Keogh, JP, Wysocki, VH, Gale, BK, Conboy, JC & Saavedra, SS 2009, 'Stable, ligand-doped, poly(bis-SorbPC) lipid bilayer arrays for protein binding and detection', ACS Applied Materials and Interfaces, vol. 1, no. 6, pp. 1310-1315. https://doi.org/10.1021/am900177p
Joubert, James R. ; Smith, Kathryn A. ; Johnson, Erin ; Keogh, John P. ; Wysocki, Vicki H. ; Gale, Bruce K. ; Conboy, John C. ; Saavedra, Steven S. / Stable, ligand-doped, poly(bis-SorbPC) lipid bilayer arrays for protein binding and detection. In: ACS Applied Materials and Interfaces. 2009 ; Vol. 1, No. 6. pp. 1310-1315.
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