Biotin-Containing Phospholipid Vesicle Layer Formed on Self-Assembled Monolayer of a Saccharide-Terminated Alkyl Disulfide for Surface Plasmon Resonance Biosensing(METHODS)
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概要
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We describe a technique to form a biotin-containing phospholipid vesicle layer on a self-assembled monolayer(SAM) deposited on a gold surface to immobilize biotinylated receptor proteins for a surface plasmon resonance(SPR) biosensor. The adsorption of vesicle of 1,2-dioleoyl-sn-glycero-3-phosphocholine(DOPC) was examined by SPR on the SAMs of dithiobis(1-deoxy-glucitol-1-carbamoyl pentane)(DDGP), 11-mercaptoundecanoic acid, 11-mercaptoundecanol, 11-amino-1-undecanethiol, and 12-mercaptododecane, and it was found that the DOPC vesicle rapidly adsorbed on the DDGP SAM to achieve the highest coverage of the surface. By quartz crystal microbalance with dissipation monitoring(QCM-D), the DOPC layer formed on the DDGP SAM was shown to be a vesicle layer, in which intact DOPC vesicles physisorbed on the SAM surface. To immobilize a biotinylated receptor protein, one of three biotinylated phospholipids, 1,2-dioleoyl-sfi-glycero-3-phosphoethanolamine-N-(biotinyl)(biotin-DOPE), N-((6-(biotinoyl)amino)hexanoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine(biotin-X-DHPE) and N-(biotmoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine(biotin-DHPE), was mixed with DOPC to form a biotin-containing vesicle layer on the DDGP SAM. A comparative binding study of NeutrAvidin and the biotin-containing vesicle layers showed that the use of biotin-X-DHPE achieved the most rapid immobilization of NeutrAvidin on the vesicle layer at the highest surface density. Furthermore, biotinylated protein A, as a receptor protein, could be immobilized through NeutrAvidin on the vesicle layer containing DOPC and biotin-X-DHPE, and its reaction with immunoglobulin G, as an analyte, was successfully observed by SPR. The results demonstrate that the biotin-containing vesicle layer on the DDGP SAM must be a useful component for SPR biosensor surfaces.
- 社団法人日本生物工学会の論文
- 2008-05-25
著者
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Morigaki Kenichi
Research Center For Environmental Genomics Kobe University:national Institute Of Advanced Industrial
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Morigaki Ken-ichi
Research Institute For Cell Engineering National Institute Of Advanced Industrial Science And Techno
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Wazawa Tetsuichi
Graduate School Of Engineering Tohoku University
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Wazawa Tetsuichi
Omron-endowed Chair In Nano Optical Devices Graduate School Of Frontier Biosciences Osaka University
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Aoyama Shigeru
Omron-endowed Chair In Nano Optical Devices Graduate School Of Frontier Biosciences Osaka University
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Ishizuka-Katsura Yoshiko
OMRON-Endowed Chair in Nano Optical Devices, Graduate School of Frontier Biosciences, Osaka Universi
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Ban Tadato
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Techn
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Ban Tadato
Research Institute For Cell Engineering National Institute Of Advanced Industrial Science And Techno
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Ishizuka-katsura Yoshiko
Omron-endowed Chair In Nano Optical Devices Graduate School Of Frontier Biosciences Osaka University
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