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    Aptamers specific for biomolecules and methods of making
    3.
    发明授权
    Aptamers specific for biomolecules and methods of making 失效
    适用于生物分子和制备方法

    公开(公告)号：US5756291A

    公开(公告)日：1998-05-26

    申请号：US484192

    申请日：1995-06-07

    申请人： Linda Griffin ,  Glenn Albrecht ,  John Latham ,  Lawrence Leung ,  Eric Vermaas ,  John J. Toole

    发明人： Linda Griffin ,  Glenn Albrecht ,  John Latham ,  Lawrence Leung ,  Eric Vermaas ,  John J. Toole

    IPC分类号： C12N9/74 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04 ,  C07K1/14

    CPC分类号： C12N9/6429 ,  C12Q1/6811 ,  C12Y304/21005

    摘要： A method for identifying oligomer sequences, optionally comprising modified base, which specifically bind target molecules such as serum proteins, kinins, eicosanoids and extracellular proteins is described. The method is used to generate aptamers that bind to serum Factor X, PDGF, FGF, ICAM, VCAM, E-selectin, thrombin, bradykinin, PGF2 and cell surface molecules. The technique involves complexation of the target molecule with a mixture of oligonucleotides containing random sequences and sequences which serve as primer for PCR under conditions wherein a complex is formed with the specifically binding sequences, but not with the other members of the oligonucleotide mixture. The complex is then separated from uncomplexed oligonucleotides and the complexed members of the oligonucleotide mixture are recovered from the separated complex using the polymerase chain reaction. The recovered oligonucleotides may be sequenced, and successive rounds of selection using complexation, separation, amplification and recovery can be employed. The oligonucleotides can be used for therapeutic and diagnostic purposes and for generating secondary aptamers.

    摘要翻译： 描述了用于鉴定特异性结合靶分子如血清蛋白，激肽，类花生酸和细胞外蛋白质的寡核苷酸序列的任选包含修饰的碱基的方法。 该方法用于产生与血清因子X，PDGF，FGF，ICAM，VCAM，E-选择蛋白，凝血酶，缓激肽，PGF2和细胞表面分子结合的适体。 该技术涉及将靶分子与含有随机序列和序列的寡核苷酸的混合物络合，所述随机序列和序列用作PCR引物的条件，其中用特异性结合序列形成复合物，但不与寡核苷酸混合物的其它成员形成。 然后将复合物与未络合的寡核苷酸分离，并使用聚合酶链反应从分离的复合物中回收寡核苷酸混合物的络合成员。 可以对回收的寡核苷酸进行测序，并且可以使用使用络合，分离，扩增和回收的连续轮次选择。 寡核苷酸可用于治疗和诊断目的并用于产生次级适体。