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1. The Significance of Biomimetic Membrane Nanobiotechnology to Biomedical Applications ......1............................. I Donald K. Martin S. I ntroduction.. ...... . . ........ ...... S.2. interaction of Lipid Membranes w ith Transport Proteins..... .......... ..... ......... .......................... ......... 3 .3. Reaction of Eukaryotic Cells Sthe Physical En iro m ent......... . .............. ........ ...... ........ 4 1.3.1. Example of the Influence of Membrane Ion Channels on the Biology of Endothelial Cells ........ 5 1.3.2. Mechanical Transduction of Stress in Lipid Bilayers ........., 8 1.4. What is the Relevance of Lipid Bilayer Membranes to N anotechnology? ........ ....... ...... ....... ............ .............................. 10 1.5. Can Biosensor Technology Benefit from Biomimetic M embrane Nanobiotechnology?i ........... ..... .............................. .... 13 1.6. Does Biomimetic Membrane Nanobiotechnology Assist in Drug Delivery? ......... ..........1. .. 15 .1 T Can Implants Benefit from Biornimetic Membrane N nnb iotechnolo gy? ..._ . ......... . . ............... .. ... 16 1.8. Concluding Remarks .. <... ...... t ... ...... . , 1........ ..... .... 17 2. Langmuir-Blodgett Technique for Synthesis of Biomimetic Lipid M embranes...... .. i . .................. ..... ... . .... 23 Agnes P. Girard-Egrot and Loic J. Blum 2.1. Introduction . ..... . ..... .... .. ........... ...... .. .... .. 23 2.21 Langmuir Monolayer Formation ....... .... ... .. _....5..... - 2,2. 1. Surface Tension,..... ...... . .... ...... ................... 26 2.2.2 . . Surfactants... ..... ... .. . ... .. . . ....... .........7.. . ....... ... . 27 2.2.3. Surface Pressure................ ...... . . 30 21214. Surface Pressure (-r) -- Area (A) Jsotherms.l..............-...... 33 2.2.5. M onolayer Stability..... .... ................... .... .. ........................ 37 2.3. Langmuir-Blodgett Technique............. . ........ 39 2.31 1. Vertical Film Deposition Principles -.............* ..,....... ..R..-... 03 2.3. L1. T. ransfer Process Energy............. - .. ..........4........... ... 41 2.3. 1 .2 Contact A ngle V alues .. ..... ................. .................... 42 2.3.1.3. Deposition Ratio......... - ,...... -. ... .... ....... .._ ,......... 43 2. 3.1.4. Advantages and Caution .. ....... ...... ......... 43 23.1 Elaboration of Organised Lipidic LB Films ................. --.... 44 2.13. Phospholipid LB Film s ....... .... ........... .......... .. ......... .... ......... 47 2.3.4. Free Supported Phospholipid LB Fil ms... ... .... ........ 5 2 .1 .. Asyminetric Phospholipid LB Bilayers ........... ................. 54 2.4. Functionalisation of Lipidic LB Filbms: Specific Features ................ 57 2.4. 1. Protein Association with the Floating Monolayer before LB Deposition ....... ...........1:;.... .57 24.12 Protein Association onto Preformed-Lipidic LB Films .......... 59 2.4.3. Oriented Protein Association in Lipidic LB Films ................. 60 .5. Trends and Prospects ..,,.... ..l. - - ....... ........ .......: 6' 3. Liposome Iechniques for Synthesis of Biomimetic Lipid rM embranes ......... ........... ... . . ..... .. ... .. 75 Stella M. Valenzuela 3. 1. Introduction .......... -- ....: . ... ... ........... .... ,. . . .. ........ ... 75 3.2. Applications and Uses of Liposomnes............................... .. ................ 75 3.3. Liposome Structure is influenced by its Phospholipid C o mi p o sitio n ... ..... ... ....... ..... .. . ...7-.. .. .... ...... .... ....... .... ... . ...... . . 76 3.4, Common Terminology Used in the Description of Liposome Structure ...... ......: rl. . ... , I...... .. .... 7...:;. 7I 3.5 Liposor e Preparation - .... .. -.... ... ....... .... ..... - . ..... .. ... . ... . .. . . . 77 3.51 . Prenaration of M ultilaimellar Vesicles.. .. .......---- ......... 78 3,5.2. Preparation of Unilamellar Vesicles.............. ..... ........ .... 79 3.5.2.1 . U itrasonication........ ........ .......... ...... .. ............. ...... 79 3 5.2.2. Extrusion through Polyiarbonate Filters .7...9.........-. 79 35.1 2.3 Freeze - Thawing...;.............. . ... 79 351214. EL thanol Injection .................. . .... .... .. 8l. 3.5.2.5 De ter ent M ethod. ... .. ... .. ...... i . . . . ..... ...... 8 1 35.2.6. Preparation of Sterile Large Unilamellar Vesicles ... 81 3.53. Preparation of Giant Uniilaellar Liposomles...... ...... . ...... 82 3.5.3.1. .Electroformation ............. ... ...........I . ....... 82 3.5.3.2. Rapid Preparation of Giant Liposomes............... 82 3.5.1. 3 Giant Unilarnamellar Liposomes Prepared in Physiologi'cal Buffeirr ....... 83 3.154. M odified Liposo es .... ...................... . ...... ........ 8 3 .5. Purifi cation of Li osom es .... ................ ............ .... ...... .. 85 4. Characterization and Analysis of Biomimetic Membranes ......... 89 Adarn 1. Mechler 4.1. Important Properties of Biomrnimetic embranes............................... 89 4.2. Methods of Characterization and Analysis .................. .................... 91 4.1. A Few Thoughts ... ........ ...... . . ......... ............... .. .. .... 9 1 4.12.2 Atomic Force Microscopy ........... .... . ........ 92 4 '. Quartz Crystal M icrobalance..... .............. .................... 96 4. 2.4. Surface Force Apparatus .... .. .. ......... .......96... ......... . 96 4,2.5. E llipsom etry ... ....97...... ........97..... .... ..... ... ..... .......... ........ ... .. 97 4.2.6. Surface P asmon Resonance ..>........... ........... -...................... 98 4.3. Coverage and M ass... . ....... .......... ........ ........... ............... ....... 99 4.4, M orphology and M echanical Properties ........1.. .................. .......... 104 4.4.1. niaging and a Few Common Artefacts ................ ............... 104 .4.2. Surface Forces and Continuum Mechanics; A FM Sim ulation .................. .. . ............................................ ..... ...107 4.4.3. M echanical Properties ........... ................. ...... .... ..... ....... ...... 118 4.5. A Brief O utlook -... ......... ........--. ... .1.... . .......-. ....... ....... -122 5. Biomimetic Membranes in Biosensor Applications.. ...... ..........127 Tfill Bcking and J. Justin Gooding 5.1. Introduction. +.. .2.7....... .. -. ...... .... 127. I;..... : ,. 5 .2 B io se n so rs .... ............. ... .. .. . .... ......... ...... ......... ........... ....... ............. ... 12 9 5 2.1 . C lasses of B iosensors..-................. ............................... ...... .. 129 5,2+2. Why Biomrnimetic Membranes for Biosensing Applications? .. 130 5.3. Biornimnetic Membranes for Biosensor Applications 1..... ............. 133 5.31. lybrid Bilayer Lipid Membranes (Supported Lipid M onolayers) ...... .... , ... .. ....... ......,......+ ,. 1... . . ... 34 5 3. 2.Solid Supported "Floatingi Bila er Lipid Membranes..... ...., 134 5,.3,.3 Tethred Bilyer l ipid Membranes. ................................... .... 137 53.3.1. Surface A ttach'ent via Low Molecular Weight 5.3.3.1 Phytanyl Lipid Derivatives for Highly nsulating M em branes ...I-. . . .. I:, i .. .... . -........ . ... .I 138 5.33,3.3 Surface Attachment via Functionalised Polymers..... 140 5.34. Lateally Structured Bilayer Lipid Membranes i.... ....,,...*+,,,. 140 5.4. Ctalytic and Affinity Biosensors Fabricated using Supported Biayer Lipid Memhbranes ...... ....... 141 5.4.1. Catalytic Biosensors based on Supported BLMs....;....-..,...., 141 5.4.2. Affinity B iosensors i........ . . .. . ........ ... ... ...... 143 5.4.2.1. Irmnunosensors based on Supported BI-Ms ..143--.i:... 143 l4.21. DNA M odified BLM s......... ......................... ............ .. 143 5.4.3. Detection of AToxins using Hybrid BLMs, Supportded BLs ad Vesices................. 143 5.4.3. General Remarks on Supported BLMs for Biosensing A pplicati ons ...... ....... .. ... ... ....... ....... .. .. .. 14 / 5.5. Membrane Biosensors Based on Ion Channel Gatinu . ... .......... 148 5.5. 1. Signal Transduction via Ion Channels,.......... . ......... 148 5.5.1. . Criteria for the Biomimetic Membrane ... .............. 148 5.5.1.2. Measurement of Membrane Conductance ........ 149 5.5.1.3. Gating of Ion Channels Incorporated into Tethered BLM s ........ .......... ....... .... . .... ...... ...... 149 5.51.4. Gating of Ion Channels Incorporated into Membranes on a Sensor Chip ....... .............. 150 5.5.2. Taking Biosensors a Step Further: The AMBRI Ion Chainel Switch Biosensor ........ ............. .... ... . 150 5.6. Concludin Remarks.............. ....... ..................... 154