Investigating Fluorescence Resonance Energy Transfer in Conjugated Liposomes

研究共轭脂质体中的荧光共振能量转移

• 批准号: 7629982
• 负责人: Punit Kohli
• 金额: $ 2.58万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-28 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7629982
• 关键词: Address; Antibodies; Antigens; Area; Biological; Biological Assay; Cells; Chemical Structure; Crosslinker; Detection; Development; Disease; Equation; Escherichia coli; Fluorescence; Fluorescence Resonance Energy Transfer; Fluorescence Spectroscopy; General Population; Genomics; Health; Imagery; Knowledge; Length; Life; Ligands; Liposomes; Measurement; Microbe; Microscopy; Molecular; Monoclonal Antibodies; NMR Spectroscopy; Optics; Performance; Phase; Process; Proteomics; Range; Research; Research Support; Solutions; Spectrum Analysis; Staphylococcus aureus; Stress; Students; Surface; System; Time; Vertebral column; Vesicle; Viola; Work; absorption; base; crosslink; design; fluorophore; improved; monomer; novel; particle; pathogen; polydiacetylene; quantum; receptor; receptor binding; response; sensor

Summary Fluorescence resonance energy transfer (FRET) has been exploited as a selective, specific, and sensitive mechanism for molecular sensing in genomics, proteomics, and other processes in living cells. For these applications, FRET relies on the remarkable efficiency of the donor-acceptor separation distance. However, to date, little research work has addressed the use of the other factors in the F¿rster equation, such as spectral overlap (J), donor-acceptor dipole orientation, and quantum yield of donor, for sensing mechanism. The overall objective of this R15 proposal application is to investigate the feasibility of using modulation of J and acceptor quantum yield (Q) as a sensing mechanism. The bilayered liposome will compose of dansyl (act as donor), polydiacetylene (PDA, act as acceptor), and receptors (such as antibodies). The interactions between receptors attached to liposomes and antigens (on the surface of a microbe) will induce stress in the conjugated PDA backbone which results in a blue spectral shift in its absorption spectrum. The consequence of PDA absorption shift results in changes in the spectral overlap (J) between donor emission and acceptor absorption which changes the FRET efficiency from donor to acceptor. The overall effect is controlled modulation of the fluorescence intensity. That is, the antibody-antigen interactions will be indirectly sensed through FRET measurements. We expect a detection limit of the proposed assay in the nanomolar range; with careful optimization, it would be possible to detect analytes in picomolar concentration range. Three specific aims for the proposed work are: (1) Design, synthesis and characterization of different diacetylene and dansyl-tagged diacetylene monomers. (2) Synthesis and characterization of fluorophores and receptors containing liposomes. (3) Detection of E. coli and S. aureus using proposed FRET-based liposome system in solution phase. Ultimately, the proposed work would provide a novel optical actuating mechanism to construct highly selective and sensitive liposomes for rapid sensing of biological molecules and particles. 1

总结 荧光共振能量转移（FRET）作为一种选择性、特异性和灵敏度高的检测方法已被广泛应用。 用于基因组学、蛋白质组学和活细胞中的其它过程中的分子传感的灵敏机制。 对于这些应用，FRET依赖于供体-受体分离的显著效率 距离然而，到目前为止，很少有研究工作涉及到F rster中其他因素的使用 计算了光谱重叠（J）、给体-受体偶极取向和给体量子产率等方程。 传感机构。本R15提案申请的总体目标是调查 利用J和受体量子产率（Q）的调制作为传感机制的可行性。 双层脂质体由丹磺酰（丹磺酰）、聚二乙炔（PDA）、聚（二苯乙炔 受体）和受体（例如抗体）。脂质体表面受体间的相互作用 和抗原（在微生物的表面上）将在缀合的PDA主链中诱导应力， 导致其吸收光谱中的蓝色光谱偏移。PDA吸收位移结果的结果 在施主发射和受主吸收之间的光谱重叠（J）的变化中，其改变了 从供体到受体的FRET效率。总的效果是荧光的受控调制 强度。也就是说，抗体-抗原相互作用将通过FRET测量来间接地感测。 我们期望所提出的检测方法的检测限在纳摩尔范围内;通过仔细优化， 将有可能检测皮摩尔浓度范围内分析物。 拟议工作的三个具体目标是： (1)不同二乙炔基和丹磺酰标记物的设计、合成与表征 二乙炔单体。 (2)含荧光团和受体的脂质体的合成与表征。 (3)E. coli和革兰氏阳性菌S.使用拟定的FRET脂质体系统 溶液相。 最后，本论文的工作将提供一种新颖的光学驱动机构来建构 用于生物分子和颗粒的快速传感。 1