Affinity-Trap Polyacrylamide Gel Electrophoresis: A Novel Method of Capturing Specific Proteins by Electro-Transfer

作者：Awada Chihiro; Sato Takashi; Takao Toshifumi^*

来源：Analytical Chemistry, 2010, 82(2): 755-761.

DOI：10.1021/ac902290q

A method for the affinity capture of specific proteins from a complex mixture using a polyacrylamide gel technique is described. The approach is based on the orthogonal electro-transfer of proteins separated by ordinary polyacrylamide gel electrophoresis (PAGE) to a ligand-coupled polyacrylamide gel (Li-PAG), which is placed under the PAGE gel. Upon electro-transfer, the proteins orthogonally migrate from the PAGE into the Li-PAG, based on the net charge. During migration to the Li-PAG, proteins that specifically interact with a ligand can be transiently trapped in the Li-PAG, while those that do not interact with a ligand pass through it. This method permits the separation of the proteins that can specifically interact with a ligand, even when present in a complex mixture. The method is demonstrated by applying it to the one-step isolation of a trypsin inhibitor from a crude extract of soybean flour.

• 出版日期2010-1-15