FSL Constructs: A Simple Method for Modifying Cell/Virion Surfaces with a Range of Biological Markers Without Affecting their Viability
FSL Constructs: A Simple Method for Modifying Cell/Virion Surfaces with a Range of Biological Markers Without Affecting their Viability
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DOI：
10.3791/3289
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发表时间：
2011-08-01
影响因子：
1.2
通讯作者：
Henry, Stephen M.
中科院分区：
文献类型：
--
作者：
Blake, Deborah A.;Bovin, Nicolai V.;Henry, Stephen M.
修饰/可视化生物表面，然后在一系列体外和体内环境中研究修饰的细胞/病毒体的能力对于进一步了解特定分子或整个实体的功能至关重要。生物表面改性的研究通常限于生物体的基因工程或化学部分与细胞表面的共价连接（1，2）。然而，这些传统技术将细胞暴露于化学反应物，或者它们需要显著的操作以实现期望的结果，使得它们繁琐，并且它们还可能无意中影响修饰细胞的活力/功能。需要一种简单的方法对细胞表面进行无害的修饰。最近，KODE技术推出了一系列由三种成分组成的新型结构：功能性头部基团（F），间隔基（S）和脂质尾部（L），称为功能间隔基脂质或FSL结构3。选择间隔物（S）以提供可分散在水中但仍会自发且稳定地掺入膜中的构建体。迄今为止，FSL构建体功能部分（F）包括一系列生物标记，包括血型相关决定簇、唾液酸、透明质酸多糖、荧光团、生物素、放射性标记和一系列肽（3-12）JSL构建体已用于修饰胚胎、精子、斑马鱼、上皮/子宫内膜细胞、红细胞和病毒体以产生质量控制系统和诊断组。修饰细胞粘附/相互作用/分离/固定，以及用于细胞/病毒体的体外和体内成像（3-12）。最常见的程序是将细胞（在无脂质培养基中）与FSL构建体的溶液在37 ℃ C4-10下孵育1-2小时。在孵育期间，FSL构建体自发地并入膜中，并且该过程完成。洗涤是可选的。被FSL构建体修饰的细胞被称为kodecytes（6-9），而病毒体是kodevirions（10）JSL构建体作为直接输注和kodecytes/kodevirions已被用于实验动物模型（7，8，10）。所有kodecytes/kodevirion似乎都保留了它们正常的活力和功能，同时获得了F部分的新功能（7，8，10，11）。FSL在生物相容性介质中的结合，自发掺入细胞膜，以及明显的低毒性，使得FSL构建了用于细胞和病毒体研究的有价值的研究工具。
The ability to modify/visualize biological surfaces, and then study the modified cell/virion in a range of in vitro and in vivo environments is essential to gaining further insight into the function of specific molecules or the entire entity. Studies of biological surface modification are generally limited to genetic engineering of the organism or the covalent attachment of chemical moieties to the cell surface(1,2). However these traditional techniques expose the cell to chemical reactants, or they require significant manipulation to achieve the desired outcome, making them cumbersome, and they may also inadvertently affect the viability/functionality of the modified cell. A simple method to harmlessly modify the surface of cells is required.Recently a new technology, KODE Technology has introduced a range of novel constructs consisting of three components: a functional head group (F), a spacer (S) and a lipid tail (L) and are known as Function-Spacer-Lipid or FSL constructs3. The spacer (S) is selected to provide a construct that is dispersible in water, yet will spontaneously and stably incorporate into a membrane. FSL construct functional moieties (F) so far include a range of saccharides including blood group-related determinants, sialic acids, hyaluronan polysaccharides, fluorophores, biotin, radiolabels, and a range of peptides(3-12).FSL constructs have been used in modifying embryos, spermatozoa, zebrafish, epithelial/endometrial cells, red blood cells, and virions to create quality controls systems and diagnostic panels, to modify cell adhesion/interaction/separation/immobilization, and for in vitro and in vivo imaging of cells/virions(3-12).The process of modifying cells/virions is generic and extremely simple. The most common procedure is incubation of cells (in lipid free media) with a solution for FSL constructs for 1-2 hours at 37 degrees C4-10. During the incubation the FSL constructs spontaneously incorporate into the membrane, and the process is complete. Washing is optional. Cells modified by FSL constructs are known as kodecytes(6-9), while virions are kodevirions(10).FSL constructs as direct infusions and kodecytes/kodevirions have been used in experimental animal models(7,8,10). All kodecytes/kodevirions appear to retain their normal vitality and functionality while gaining the new function of the F moiety(7,8,10,11).The combination of dispersibility in biocompatible media, spontaneous incorporation into cell membranes, and apparent low toxicity, makes FSL constructs valuable research tools for the study of cells and virions.