发明授权
- 专利标题: Aptamers specific for biomolecules and methods of making
- 专利标题(中): 适用于生物分子和制备方法
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申请号: US484192申请日: 1995-06-07
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公开(公告)号: US5756291A公开(公告)日: 1998-05-26
- 发明人: Linda Griffin , Glenn Albrecht , John Latham , Lawrence Leung , Eric Vermaas , John J. Toole
- 申请人: Linda Griffin , Glenn Albrecht , John Latham , Lawrence Leung , Eric Vermaas , John J. Toole
- 申请人地址: CA Foster City
- 专利权人: Gilead Sciences, Inc.
- 当前专利权人: Gilead Sciences, Inc.
- 当前专利权人地址: CA Foster City
- 主分类号: C12N9/74
- IPC分类号: C12N9/74 ; C12Q1/68 ; C07H21/02 ; C07H21/04 ; C07K1/14
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.
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