公开(公告)号：US06447764B1

公开(公告)日：2002-09-10

申请号：US09505963

申请日：2000-02-17

申请人： Ernst Bayer ,  Hans Fritz ,  Martin Maier ,  Jens Schewitz ,  Michael Gerster

发明人： Ernst Bayer ,  Hans Fritz ,  Martin Maier ,  Jens Schewitz ,  Michael Gerster

IPC分类号： A61K3174

CPC分类号： B01J41/04 ,  C12N15/1006 ,  C12Q1/6806 ,  Y10S977/906

摘要： The invention relates to a method for isolating anionic organic substances from aqueous systems using polymer nanoparticles with cationic pH-sensitive surface groups. Extraction can be carried out directly from diluted solutions, biological media (blood plasma, serum, urine, etc.) and complex buffer systems (e.g. PCR preparations contain detergents) without prior derivatization of the samples or addition of binding buffers. After separation from the surrounding medium the conjugates of the corresponding substances and polymer nanoparticles obtained in this manner can be purified through additional washing steps and effectively desalinated. Owing to the pH-sensitivity of the basic surface groups, the bound substances can be released in a targeted manner after separation by modification of the pH of the medium. By using volatile bases contamination of the released samples with ionic residues can be avoided. By adding small quantities of SDS and acetonitrile during the separation stage, the sensitivity of the method can be raised further, especially when the substance to be isolated are present in low concentrations (

摘要翻译： 本发明涉及使用具有阳离子pH敏感表面基团的聚合物纳米颗粒从水性体系中分离阴离子有机物质的方法。 提取可以直接从稀释溶液，生物培养基（血浆，血清，尿液等）和复合缓冲液系统（例如PCR制剂含有洗涤剂）进行，而无需样品的先前衍生或添加结合缓冲液。 在与周围介质分离后，可以通过另外的洗涤步骤纯化相应物质和以这种方式获得的聚合物纳米颗粒的共轭物，并有效脱盐。 由于碱性表面基团的pH敏感性，可以通过改变介质的pH进行分离后以目标方式释放结合物质。 通过使用挥发性碱可以避免释放的样品与离子残留物的污染。 通过在分离阶段添加少量的SDS和乙腈，可以进一步提高该方法的灵敏度，特别是当待分离的物质以低浓度（<1mumol / L）存在时。 该方法是通用应用的一种提取方法，适用于低分子阴离子化合物以及肽，核酸和核酸衍生物。 恢复率高达100％。