公开(公告)号：US06881540B2

公开(公告)日：2005-04-19

申请号：US09791317

申请日：2001-02-23

申请人： Steven Cramer ,  Kaushal Rege ,  Jonathan Dordick

发明人： Steven Cramer ,  Kaushal Rege ,  Jonathan Dordick

IPC分类号： B01D15/42 ,  C07K1/22 ,  C12Q1/68

CPC分类号： C07K1/22 ,  B01D15/422

摘要： An effective technique for the high throughput screening of displacers is described. In this technique, potential displacers are employed to displace a biomolecule (e.g., protein) adsorbed on a chromatographic resin in small-scale batch displacement experiments. The amount of protein displaced from a specific resin by a defined concentration of displacer is determined by monitoring the supermatant for the protein. By evaluating the displaced protein rather than the displacer itself, this technique enables a single detection technique (e.g., absorbance, fluorescence, etc.) to be employed for all batch displacement experiments. By monitoring the amount of protein displaced, the effacy of a large number of potential displacers can be rapidly evaluated. The entire experimental procedure can be carried out rapidly and is thus amenable to high throughput parallel screening of molecules possessing a large range of affinities and physico-chemical properties. The error of the technique is within 5% of protein displaced, thus making it a very reliable technique. The technique can be extended to different stationary phase materials, biomolecules, and modes of interaction.

摘要翻译： 描述了用于置换器的高通量筛选的有效技术。 在该技术中，使用潜在的置换剂来置换小规模批量置换实验中吸附在色谱树脂上的生物分子（例如蛋白质）。 通过监测蛋白质的超级蛋白质来确定由特定树脂从限定浓度的置换剂中排出的蛋白质的量。 通过评估置换的蛋白质而不是置换剂本身，该技术使得能够将单个检测技术（例如，吸光度，荧光等）用于所有的批量置换实验。 通过监测蛋白质置换量，可以快速评估大量潜在置换剂的效率。 整个实验程序可以快速进行，因此适合高通量平行筛选具有大范围亲和力和物理化学性质的分子。 该技术的误差在蛋白质置换的5％以内，因此使其成为非常可靠的技术。 该技术可以扩展到不同的固定相材料，生物分子和相互作用模式。