Moaddel Ruin, Wainer Irving W
Laboratory of Clinical Investigation, Gerontology Research Center, National Institute on Aging/NIH, 5600 Nathan Shock Drive, Baltimore, Maryland 21224, USA.
Nat Protoc. 2009;4(2):197-205. doi: 10.1038/nprot.2008.225.
Cellular membrane affinity chromatography is a technique that is based on the immobilization of a target trans-membrane protein onto a stationary phase. The target protein is isolated by homogenization and solubilization of a source (e.g., cell line) followed by immobilization on either the immobilized artificial membrane-phosphatidyl choline (IAM-PC) stationary phase or the surface of an open tubular capillary during a dialysis step. The procedure typically takes 3-4 d for the IAM-PC stationary phase, whereas the open-tubular method takes an extra week for the preparation of the capillary. The resulting columns can then be used to characterize binding sites on the target protein through frontal chromatographic and/or nonlinear chromatographic studies using a wide variety of ligands including small molecules and polypeptides. The columns have been used in drug discovery as well as in the screening of tobacco smoke condensates.
细胞膜亲和色谱法是一种基于将目标跨膜蛋白固定在固定相上的技术。通过对来源（如细胞系）进行匀浆和溶解，然后在透析步骤中将其固定在固定化人工膜 - 磷脂酰胆碱（IAM - PC）固定相或开放管状毛细管表面，从而分离出目标蛋白。对于IAM - PC固定相，该过程通常需要3 - 4天，而开放管状方法则需要额外一周来制备毛细管。然后，所得的色谱柱可用于通过使用包括小分子和多肽在内的多种配体进行前沿色谱和/或非线性色谱研究来表征目标蛋白上的结合位点。这些色谱柱已用于药物发现以及烟草烟雾冷凝物的筛选。
