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    • 1. 发明授权
    • High surface density covalent immobilization of oligonucleotide monolayers using a 1-(thiotrifluoroacetato)-11-(trichlorososilyl)-undecane linker
    • 使用1-(三(三氟乙酰))-11-(三氯硅烷基) - 十一烷接头的高密度共价固定寡核苷酸单层
    • US06169194A
    • 2001-01-02
    • US08951448
    • 1997-10-16
    • Michael ThompsonMark E. McGovern
    • Michael ThompsonMark E. McGovern
    • C07F712
    • B82Y30/00B01J20/3242B32B17/10688B82Y5/00B82Y15/00C03C17/30C03C17/3405C07B2200/11C07H21/00Y10S977/702Y10S977/902Y10S977/92Y10S977/924Y10S977/958
    • Oligonucleotides and other biomolecules are immobilized in high density on solid substrates through covalent forces using either a permanent thioether bond, or a chemoselectively reversible disulfide bond to a surface thiol. Substrates which have hydroxyl groups on their surfaces can be first silanized with a trichlorosilane containing 2-20 carbon atoms in its hydrocarbon backbone, terminating in a protected thiol group. The oligonucleotides or other biomolecules are first connected to a tether consisting of a hydrocarbon or polyether chain of 2-20 units in length which terminates in a thiol group. This thiol may be further modified with a halobenzylic-bifunctional water soluble reagent which allows the conjugate to be immobilized onto the surface thiol group by a permanent thioether bond. Alternatively, the oligonucleotide-tether-thiol group can be converted to a pyridyldisulfide functionality which attaches to the surface thiol by a chemoselectively reversible disulfide bond. The permanently bound oligonucleotides are immobilized in high density compared to other types of thiol functionalized silane surfaces and to the avidin-biotin method.
    • 寡核苷酸和其他生物分子通过共价力使用永久硫醚键或与表面硫醇的化学选择性可逆二硫键以高密度固定在固体基质上。 在其表面上具有羟基的底物可以首先在其烃骨架中用含有2-20个碳原子的三氯硅烷硅烷化,终止于受保护的硫醇基团。 寡核苷酸或其他生物分子首先连接到由长度为2-20单位的烃或聚醚链组成的系链,其终止于硫醇基团。 该硫醇可以用卤代苄基双功能水溶性试剂进一步改性,其允许缀合物通过永久硫醚键固定在表面硫醇基上。 或者,寡核苷酸 - 系链 - 硫醇基团可以转化成通过化学选择性可逆的二硫键附着于表面硫醇的吡啶基二硫键官能团。 与其他类型的硫醇官能化硅烷表面和抗生物素蛋白 - 生物素方法相比,永久结合的寡核苷酸以高密度固定。