Stable, ligand-doped, poly(bis-SorbPC) lipid bilayer arrays for protein binding and detection.
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 GM(1), and the extent of binding was correlated to the mole percentage of GM(1) in each spot. A poly(lipid) bilayer array composed of spots doped with GM(1) 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)/GM(1) 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.
Joubert
James R
JR
Department of Chemistry, University of Arizona, 1306 E. University Boulevard, Tucson, Arizona 85721, USA.
Smith
Kathryn A
KA
Johnson
Erin
E
Keogh
John P
JP
Wysocki
Vicki H
VH
Gale
Bruce K
BK
Conboy
John C
JC
Saavedra
S Scott
SS
D000327:Adsorption; D008024:Ligands; D008051:Lipid Bilayers / Q000737:chemistry*; D008422:Materials Testing; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*
D008024:Ligands; D008051:Lipid Bilayers; D011506:Proteins