Patent ID: 7825227

Claim:
A method for purifying a protein complex and individual components comprising the complex from a cell, a cell or tissue lysate, or an organism, comprising the steps carried out in the order set forth of: a) providing a first binding component comprising four parts: 1) first affinity tag, 2) a protease specificity segment, 3) second affinity tag, and 4) a bait, wherein the first affinity tag has a greater affinity for its binding partner than the second affinity tag has for its binding partner, wherein the first affinity tag is biotinylated biotinylation recognition site, wherein said recognition sequence comprises the sequence set forth in SEQ ID NO:6, wherein the second affinity tag is proximal to the bait, and wherein said second affinity tag is His 6 hexapeptide, and wherein the protease specificity segment is located between said first affinity tag and said second affinity tag; b) contacting the first binding component with a component of the protein complex, whereby part 4) of the first binding component binds to said component of the protein complex, thereby forming a bait-bound protein complex; c) contacting the complex formed in step b) with a first affinity matrix specific for the first affinity tag thereby binding said complex to said matrix, and separating the complex from unbound material, and; d) separating the individual components of the complex from the first binding component bound to the matrix, whereby the purified individual components are obtained.