本研究以紅外光譜法探討白蛋白對於陰陽離子液胞雙層膜中分子碳氫鏈排列的影響，以及添加之膽固醇在其中所扮演的角色。首先由陽離子型界面活性劑hexadecyltrimethylammonium bromide（HTMAB）與陰離子型界面活性劑sodium dodecylsulfate（SDS）製備出離子對雙親分子（ion pair amphiphile） hexadecyltrimethylammonium-dodecylsulfate （HTMA-DS）。然後將HTMA-DS與雙碳氫鏈二甲基溴化銨（dialkyldimethylammonium bromide, DXDAB）以等莫耳比例混合，並利用適當製程製備出帶正電的陰陽離子液胞(catanionic vesicle)。
實驗結果顯示在HTMA-DS/DXDAB所形成的液胞雙層膜中，分子碳氫鏈都呈現有序的排列，但含雙十四烷基二甲基溴化銨（ditetradecyldimethylammonium bromide, DTDAB）之液胞的物理穩定性顯得較差。在HTMA-DS/DXDAB之液胞分散液中加入白蛋白後，發現白蛋白的吸附會造成液胞界面電位的下降，且白蛋白濃度較高時，液胞中分子碳氫鏈的排列變得較鬆散。其中以含DTDAB之液胞的物理穩定性及分子碳氫鏈排列，受到白蛋白的影響較大。在莫耳比1/1之HTMA-DS/DXDAB的液胞中添加膽固醇時，隨著膽固醇含量增加，液胞雙層膜中分子碳氫鏈的排列顯得越鬆散，且液胞的界面電位變得越高。在高濃度白蛋白存在下，液胞雙層膜中分子碳氫鏈的排列則變得更鬆散。相較於未含膽固醇之液胞的雙層膜，含膽固醇之液胞雙層膜中的分子碳氫鏈排列受到白蛋白的影響更大。
在HTMA-DS/DXDAB之液胞系統中，含DTDAB之液胞的物理穩定性明顯較差，可能是因為HTMA-DS與DTDAB在雙層膜中的相容性不佳，所以液胞易受到白蛋白破壞。添加膽固醇至液胞中，雖然液胞雙層膜中分子碳氫鏈排列變得鬆散，白蛋白似乎易與雙層膜中的分子碳氫鏈發生疏水作用，但膽固醇可能改善了HTMA-DS與DTDAB之間的相容性，因此提升了液胞在白蛋白存在下的物理穩定性。

This study investigated the effects of bovine serum albumin (BSA) on the alkyl chain arrangement in bilayers of catanionic vesicles and the role of added cholesterol in the effects by fourier transform infrared (FTIR) spectroscopy. The ion pair amphiphile, hexadecyltrimethylammonium- dodecylsulfate (HTMA-DS), was first prepared from the cationic surfactant, hexadecyltrimethylammonium bromide (HTMAB), and the anionic surfactant, sodium dodecylsulfate (SDS). HTMA-DS was then mixed with dialkyldimethylammonium bromide (DXDAB) with 1:1 molar ratio to form positively charged catanionic vesicles by an appropriate process.
The experimental results indicated that the arrangement of molecular alkyl chains was ordered in the bilayers of HTMA-DS/DXDAB vesicles. However, the physical stability of HTMA-DS/ ditetradecyldimethylammonium bromide (DTDAB) vesicles was apparently lower. It was found that the zeta potential of vesicles was decreased by the BSA adsorption after adding BSA into the dispersion of HTMA-DS/DXDAB vesicles. In addition, the arrangement of the molecular alkyl chains in the vesicles became disordered with the presence of high concentrations of BSA. The physical stability and arrangement of the molecular alkyl chains of vesicles containing DTDAB were especially affected by BSA. By adding of cholesterol into the vesicles of HTMA-DS/DXDAB with a molar ratio of 1:1, one could find that the arrangement of the alkyl chains in the vesicular bilayers became disordered and the zeta potential of the vesicles was increased. With the presence of high concentrations of BSA, the arrangement of molecular alkyl chains in the vesicular bilayers became further less packed. In comparison to the vesicles without containing cholesterol, the alkyl chain arrangement of vesicles containing cholesterol was much affected by BSA.
In the HTMA-DS/DXDAB vesicle systems, lower physical stability was detected for the vesicles containing DTDAB, which was probably due to the poor compatibility of HTMA-DS and DTDAB in the bilayers. The vesicles were thus easily disrupted by BSA. With the addition of cholesterol into the vesicles, though the vesicular bilayers became less packed and BSA seemed easily interacted with the molecular alkyl chains in the bilayers through the hydrophobic effects, the compatibility of HTMA-DS and DTDAB might be improved by cholesterol and thus the physical stability of the vesicles in the presence of BSA was enhanced.

Bach, D., and Wachtel, E., “Phospholipid/cholesterol model membranes: formation of cholesterol crystallites,” Biochim. Biophys. Acta 1610, 187-197, 2003.

Byler, D. M., Susi, H., and Regional, E., “Examination of the secondary structure of proteins by deconvolved FTIR Spectra ,” Biopolymers 25, 469-487, 1986.

Carrion, F. J., Maza, A. D., and Parra, J. L., “The influence of ionic strength and lipid bilayer charge on the stability of liposomes,” Colloid. Interf. Sci. 164, 78-87, 1994.

Carvalho, L. A., and Carmona-Ribeiro, A. M., “Interaction between cationic vesicles and serum proteins, ” Langmuir 14, 6077-6081, 1998.

Charbonneau, D., Beauregard, M., and Tajmir-Riahi, H., “Structural analysis of human serum albumin complexes with cationic lipids,” J. Phys. Chem. B 113, 1777-1784, 2009.

Charbonneau, D., and Tajmir-Riahi, H., “Study on the interaction of cationic lipids with bovine serum albumin,” J. Phys. Chem. B 114, 1148-1155, 2010.

Choosakoonkriang, S., Wiethoff, C. M., Anchordoquy, T. J., Koe, G. S., Smith, J. G., and Middaugh, C. R., “Infrared spectroscopic characterization of the interaction of cationic lipids with plasmid DNA”, the journal of biological chemistry 276, 8037-8043, 2001.

Ciani, L., Ristori, S., Salvati, A., Calamai, L., Martini, G., “DOTAP/DOPE and DC-CHOL/DOPE lipoplexes for gene delivery: zeta potential measurements and electron spin resonance spectra,” Biochim. Biophys. Acta 1664, 70-79, 2004

Dhanikula , A. B., and Panchagnula, R., “Fluorescence anisotropy, FT-IR spectroscopy and 31-P NMR studies on the interaction of paclitaxel with lipid bilayers,” Lipids 43, 569-579, 2008.

Dong, A., Huang, P., and Caughey, W. S., “Protein secondary structures in water from second-derivative amide I infrared spectra,” Biochemistry 29, 3303-3308, 1990.

Du, N., Song, S. E., and Hou, W. G., “Vesicle stability in aqueous mixtures of zwitterionic/anionic surfactants,” Colloids Surf. A: Physicochem. Eng. Aspects 312, 104-112, 2008.

Farhood, H., Serbina, N., and Huang, L., “The role of dioleoyl phophatidylethanolamine in carionic liposome mediated gene transfer,” Biochim. Biophys. Acta, 1235, 289-295,1995.

Feitosa, E., Jansson, J., and Lindman, B., “The effect of chain length on the melting temperature and size of dialkyldimethylammonium bromide vesicles,” Chemi. Phys. Lipids 142, 128-132, 2006

Fournier, I., Barwicz, J., Auger, M., and Tancrède, P., “The chain conformational order of ergosterol- or cholesterol-containing DPPC bilayers as modulated by Amphotericin B: a FTIR study,” Chemis. Phys. Lipids 151, 41-50, 2008.

Frailea, M. V., Blanco-Melgara, M., Martínez, M. R., López-Rodrígueza , G., Gallego-Nicasioa, J. A., and Carmonab, P., “Structure and interactions of albumin-lipid systems as studied by infrared spectroscopy,” J. Molecular Structure 651-653, 231-236, 2003.

Israelachvili, J. N., Mitchell, D. J., and Ninham, B. W., “Theory of self-assembly of hydrocarbon amphiphiles into micelles and bilayers,” J. the Chemi. Soc.-Faraday Transa. 72, 15-25, 1976.

Kaler, E. W., Murthy, A. K., Rodriguez, B. E., and Zasadzinski, A. N.,“Spontaneous vesicle formation in aqueous mixtures of single-tailedsurfactants,” Science 245, 1371-1374, 1989.

Kamilya, T., Pal, P., Mahato, M., and Talapatra, G. B., “Immobilization and the conformational study of phospholipid and phospholipid-protein vesicles,” Materials Sci. Eng. C 29 , 1480-1485, 2009.

Kodati, V. R and Lafleur, M., “Comparison between orientational and conformational orders in fluid lipid bilayers,” Biophys. J. 64, 163-170, 1993.

Kodati, V. R., El-Jastimi, R., and Lafleur, M., “Contribution of the intermolecular coupling and librotorsional mobility in the methylene stretching modes in the infrared spectra of acyl chains,” J. Phys. Chem. 98, 12191-12197, 1994.

Lasic, D. D., “Liposomes: from physics to applications,” Elsevier, New York, 265-318, 1993.

Lewis, R. N.A.H., and McElhaney, R. N., “The structure amd organization of phospholipid bilayers as revealed by infrared spectroscopy,” Chem. Phys. Lipids 96, 9-21, 1998.

Liu, D. Z., Chen W. Y., .Tasi, L. M., and Yang, S.P., “Microcalorimetric and shear studies on the effects of cholesterol on the physical stability of lipid vesicles,” Colloids Surf. A: Physicochem. Eng. Aspects 172, 57-67, 2000.

Manosroi A., Wongtrakul, P., Manosroi, J., Sakai, H., Sugawara, F., Yuasa, M.,and Abe, M., “Characterization of vesicles prepared with various non-ionic surfactants mixed with cholesterol,” Colloids Surf. B: Biointerfaces 30, 129-138, 2003.

Matsumura, H., Watanabe, K., and Furusawa, K., “Flocculation behavior of egg phosphatidylcholine liposomes caused by Ca 2 + ions,” Colloids Surf. A: Physicochem. Eng. Aspects 98, 175-184, 1998.

McMullen, T. P. W., Lewis, N. A. H., and McElhaney, R. N., “Differential scanning calorimetric study of the effect of cholesterol on the thermotropic phase behavior of a homologous series of linear saturated phosphatidylcholines,” Biochemistry 32, 516-522, 1993.

Meierhofer, T., Elsen, J. M. H., Cameron, P. J., Muñoz-Berbel , X., Jenkins, A.,and Toby. A., “The interaction of serum albumin with cholesterol containing lipid vesicles,” J. Fluoresc. 20, 371-376, 2010.

Mel’nikova, Y. S., Mel’nikov, S. M., and Lofroth, J. E., “Physico-chemicalaspects of the interaction between DNA and oppositely charged mixed liposomes,” Biophysical Chemistry 81, 125-141, 1999.

New, R. R. C., “Liposomes: a practical approach,” Oxford University Press, New York, 1-32, 1990.

Ohtsuka, I., and Yokohama, S., “Penetration of bovine serum albumin into dipalmitoylphosphatidylglycerol monolayers: direct observation by atomic force microscopy,” Chem. Pharm. Bull. 53, 42-47 , 2005.

Regev, O., and Khan, A., “Alkyl chain symmetry effects in mixed cationic-anionic surfactant systems,” Colloid Interf. Sci. 182, 95-109, 1996.

Róg, T., Pasenkiewicz-Gierula, M., Vattulainen, I., and Karttunen, M., “Ordering effects of cholesterol and its analogues,” Biochim. Biophys. Acta 1788 , 97-121, 2009.

Semple, S. C., Chonn, A., and Cullis, P. R., “Influence of cholesterol on the association of plasma proteins with liposomes,” Biochemistry 35, 2521-2525, 1996.

Silvius, J. R., Giudice , D., and Lafleur, M., “Cholesterol at different bilayer concentrations can promote or antagonize lateral segregation of phospholipids of differing acyl chain length,” Biochemistry, 35, 15198-15208, 1996.

Taira, M. C., Stern, A. L., Escobar, M. E., Adragna, N., and Alonso-Romanowski, S., “Interaction of sodium nitroprusside/liposome system with bovine serum albumin: a short-cut determination,” Biotechnology Letters 25, 1425-1429, 2003.

Tondre, C., and Caillet, C., “Properties of the amphiphilic films in mixed cationic/anionic vesicles: a comprehensive view from a literature analysis,”Adv. Colloi. Interfa. Sci. 93, 115-134, 2001.

Tsunoda, T., Imura, T., Kadota, M., Yamazaki, T., Yamauchi, H., Kwon, K. O., Yokoyama, S., Sakai, H., and Abe, M., “Effect of lysozyme and bovine serum albumin on membrane characteristics of dipalmitoylphosphatidylglycerol liposomes,” Colloids. Surf. B: Biointerfaces 20, 155-163, 2001.

Venkatatraman, N. V., and Vasudevan, S., “Cholesterol binding to the alkyl chains of an intercalated surfactant bilayer,” J. Phys. Chem. B 107, 10119-10126, 2003.

Vist M. R., and Davis J, H., “Phase equilibria of cholesterol/ dipalmitoylphosphatidylcholine mixtures: 2HNuclear magnetic resonance and differential scanning calorimetry,” Biochemistry 29, 451-464, 1990.

Yokouchi, Y., Tsunoda, T., Imura, T., Yamauchi, H., Yokoyama, S., Sakai, H., and Abe, M., “Effect of adsorption of bovine serum albumin on liposomal membrane characteristics,” Colloids Surfa. B: Biointerfaces 20, 95-103, 2001.

Zimmerman , R. J., Kanal , K. M., Sanders, J., Cameron, I. L., and Fullerton, G. D., “Osmotic pressure method to measure salt induced folding/unfolding of bovine serum albumin,” J. Biochem. Biophys. Methods 30 , 113-131, 1995.

呂奇達，“帶正電的陰陽離子液胞與DNA 之結合行為的探討，”國立成功大學化學工程學系碩士論文，2005。

李威漢，“陰陽離子界面活性劑的製備及其相轉移行為的熱卡分析，” 國立成功大學化學工程學系碩士論文，2010。

林冠豪，“帶電的陰陽離子液胞之製備及物理穩定性研究，”國立成功大學化學工程學系碩士論文，2004。

陳柏瑋，“白蛋白存在下帶正電陰陽離子液胞穩定性的探討，”國立成功大學化學工程學系碩士論文，2007。

洪振益，“溫度效應對帶電陰陽離子液胞釋放行為的影響，”國立成功大學化學工程學系碩士論文，2009。

黃鉦琳，“帶電陰陽離子液胞的形成及其膠化之研究，”國立成功大學化學工程學系碩士論文，2007。

德芮克國際股份有限公司，“Nano ZS 雷射光散射法粒徑及界面電位量測儀儀器相關說明，”台南，2005。

鍾依玲，“陰/陽離子液胞自發性形成之探討，”國立成功大學化學工程學系碩士論文，2002。