Kirkegaard L, Agee C C
Proc Natl Acad Sci U S A. 1973 Aug;70(8):2424-8. doi: 10.1073/pnas.70.8.2424.
Intervent dilution chromatography separates interacting macromolecules by subjecting them to a dynamic environment in which the association constant is continuously varied. The dynamic environment is produced by using the sieving properties of a gel to repeatedly propel the molecular complex across an intervent boundary. Behind the boundary, at high intervent concentrations, the complex dissociates; ahead of the boundary, the component molecules are separated by adsorption processes. By selective adjustment of the intervent composition of the sample and the conditions of column equilibration, the process is adapted to a particular need. This report describes the chromatographic concept and shows how parameters are adjusted to obtain the desired separation. In this study a particularly difficult separation, i.e., the separation of ribosomal proteins from ribosomal RNA, is chosen to illustrate the power of the procedure.
介入稀释色谱法通过使相互作用的大分子处于缔合常数不断变化的动态环境中来分离它们。这种动态环境是利用凝胶的筛分特性反复推动分子复合物穿过介入边界而产生的。在边界之后，介入剂浓度较高时，复合物解离；在边界之前，组分分子通过吸附过程分离。通过选择性调节样品的介入剂组成和柱平衡条件，该过程可适应特定需求。本报告描述了色谱概念，并展示了如何调整参数以实现所需的分离。在本研究中，选择了一种特别困难的分离，即从核糖体RNA中分离核糖体蛋白，以说明该方法的功效。
