Method and apparatus for degradation and sequencing of polymers which sequentially eliminate terminal residues
申请人:PERSEPTIVE BIOSYSTEMS, INC.
公开号:EP0639607A2
公开(公告)日:1995-02-22
A method and apparatus for sequentially degrading at least a portion of a polymer of backbone repeating units, the polymer having a terminal repeating unit comprised of a nucleophile and a backbone carbonyl carbon distant from the nucleophile, comprising the steps of first initiating attack of said nucleophile upon said backbone carbonyl carbon by raising the energy level to activate said nucleophile for said attack. Secondly, forming a ring comprising the terminal repeating unit, thereby simultaneously releasing the ring and generating a shortened polymer having a terminal repeating unit capable of nucleophile attack upon the backbone carbonyl carbon and, lastly, maintaining the reaction conditions necessary for repeating steps a and b until the portion of the polymer desired is degraded. In a related embodiment, polyamide nucleic acid (PNA) sequences can be determined by generating a nested set of polymer fragments, each fragment having N-x repeating units where N is the total number of repeating units in the parent polymer and x is the number of degradation cycles the fragment has been subjected to, and then analyzing the nested set of polymer fragments to determine polymer sequence. An apparatus embodying the method of sequential degradation is also described. Analysis may be by MALD-TOF.
一种顺序降解至少部分骨架重复单元聚合物的方法和装置,该聚合物具有由亲核物和远离亲核物的骨架羰基碳组成的末端重复单元,包括以下步骤:首先,通过提高能级以激活所述亲核物对所述骨架羰基碳的攻击,启动所述亲核物对所述骨架羰基碳的攻击。其次,形成一个包含末端重复单元的环,从而同时释放该环并生成一个缩短的聚合物,其末端重复单元能够对骨架羰基碳进行亲核攻击;最后,保持重复步骤 a 和 b 所需的反应条件,直到所需的聚合物部分降解为止。在一个相关的实施方案中,聚酰胺核酸(PNA)序列可以通过生成嵌套的聚合物片段来确定,每个片段具有 N-x 个重复单元,其中 N 是母体聚合物中重复单元的总数,x 是片段所经历的降解循环次数,然后分析嵌套的聚合物片段来确定聚合物序列。此外,还介绍了一种体现顺序降解方法的装置。分析可采用 MALD-TOF。