PMID- 19178341
OWN - NLM
STAT- MEDLINE
DCOM- 20090304
LR  - 20090130
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 81
IP  - 3
DP  - 2009 Feb 1
TI  - Regenerable tethered bilayer lipid membrane arrays for multiplexed label-free 
      analysis of lipid-protein interactions on poly(dimethylsiloxane) microchips using 
      SPR imaging.
PG  - 1146-53
LID - 10.1021/ac8023137 [doi]
AB  - We report a microfabrication approach to generate well-defined, addressable, and 
      regenerable lipid membrane arrays in poly(dimethylsiloxane) (PDMS) microchips for 
      label-free analysis of lipid-protein interactions with surface plasmon resonance 
      imaging (SPRi). The multiplexed detection is demonstrated with a tethered bilayer 
      membrane array built in parallel microchannels. These channels allow multiple 
      measurements to be carried out simultaneously, showing low deviations for 
      element-to-element variation in quantifiable signal. Lipid-conjugated receptors 
      were utilized as model systems for protein binding analysis, and the feasibility 
      of regenerating the tethering sublayer after binding was investigated. The 
      results show that the lipid membrane can be removed effectively by nonionic 
      surfactant Triton X-100. The small variance in SPR signal for the buildup 
      process, i.e., <4% RSD for 3 cycles of detection, removal, and regeneration, 
      indicates the sensing interface is highly reproducible. A calibration curve was 
      obtained for cholera toxin using the monosialoganglioside (GM1) receptor, 
      displaying a linear relationship in the 25 to 175 microg/mL range with a limit of 
      detection of 260 nM. In addition, interaction of a phosphatidylinositol (PIP) 
      with its binding protein and biotin/avidin interactions were employed for array 
      measurements. To further enhance the SPR detection signal, a layer-by-layer 
      amplification strategy was demonstrated that uses biotinylated antibody, 
      NeutrAvidin and biotinylated anti-avidin, and the signal for protein binding on 
      the membrane increased by 400%. The tethered membrane array technology, in 
      combination with SPRi, offers an attractive platform for studies of membrane 
      proteins, and can also find a range of applications for rapid screening of drug 
      candidates interacting with proteins embedded in the near-native environment.
FAU - Taylor, Joseph D
AU  - Taylor JD
AD  - Department of Chemistry, University of California, Riverside, California 92521, 
      USA.
FAU - Linman, Matthew J
AU  - Linman MJ
FAU - Wilkop, Thomas
AU  - Wilkop T
FAU - Cheng, Quan
AU  - Cheng Q
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Dimethylpolysiloxanes)
RN  - 0 (Lipid Bilayers)
RN  - 0 (Lipids)
RN  - 0 (Membrane Proteins)
RN  - 63148-62-9 (baysilon)
SB  - IM
MH  - Dimethylpolysiloxanes/*chemistry
MH  - Lipid Bilayers/*chemistry
MH  - Lipids/chemistry
MH  - Membrane Proteins/*chemistry
MH  - Microfluidic Analytical Techniques/*methods
MH  - Surface Plasmon Resonance/*methods
EDAT- 2009/01/31 09:00
MHDA- 2009/03/05 09:00
CRDT- 2009/01/31 09:00
PHST- 2009/01/31 09:00 [entrez]
PHST- 2009/01/31 09:00 [pubmed]
PHST- 2009/03/05 09:00 [medline]
AID - 10.1021/ac8023137 [pii]
AID - 10.1021/ac8023137 [doi]
PST - ppublish
SO  - Anal Chem. 2009 Feb 1;81(3):1146-53. doi: 10.1021/ac8023137.