Participants/Project Abstracts
Summer 2007 Bioanalytical Science REU Participants
CHARACTERIZATION OF CHEMICAL STRUCTURE OF A REGENERABLE TETHERED BILAYER MEMBRANE USING A"PEPTIDE CUSHION" ON AN SPR SUBSTRATE
Owen
E. Holder, Joseph Taylor, and Dr. Quan ‘Jason’ Cheng
Department
of Chemistry, University of California—Riverside, UCR 92521 and
Vanderbilt University, Nashville, TN, 37235
Abstract
In this study, surface plasmon resonance (SPR)
spectroscopy is used as a novel means to elucidate structural information about
tethered bilayer lipid membranes. The tethered membrane is generated by
sequentially layering biotinylated bovine serum albumin directly on gold, then
a linking layer of avidin for capture of biotinyated unilamellar vesicles,
followed by promoted fusion using a concentrated solution of PEG-8000. SPR is
advantageous for providing both label-free detection and real-time observation
in a flow injection biosensing device. However, a universal drawback in SPR
deals with the expensive gold substrates employed, which are limited to
one-time-use in the apparatus. Here, we have discovered a simple method to
circumvent this shortcoming by developing a regenerable bilayer membrane. This
technique enables us to execute multiple experiments on a single gold chip
simply using a low concentration of the nonionic surfactant Triton X-100 to
remove the lipids from the protein sublayers. This procedure drastically cuts
down on experimental time, setup of the substrates, as well as cost. In
addition, our research may shed light on the unique characteristics of avidin
protein, in which we have utilized its unique binding with biotin to form a
"peptide cushion" for lipidic structures. Future plans aim at developing this
technique for SPR imaging, where this thin-film regeneration technique would be
directed towards membrane arrays constructed within a microfluidic device
capable of performing multiple experiments on a single substrate.
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