US 12,358,947 B2
Method for automated protein purification
Ryan Stikeleather, Phoenix, AZ (US)
Assigned to Arizona Board of Regents on Behalf of Arizona State University, Scottsdale, AZ (US)
Filed by Ryan Stikeleather, Phoenix, AZ (US)
Filed on Apr. 28, 2021, as Appl. No. 17/242,973.
Claims priority of provisional application 63/024,050, filed on May 13, 2020.
Prior Publication US 2021/0355161 A1, Nov. 18, 2021
Int. Cl. C07K 1/36 (2006.01); C07K 1/22 (2006.01)
CPC C07K 1/36 (2013.01) [C07K 1/22 (2013.01)] 10 Claims
 
1. A method for purification of a target protein fused to an affinity tag, comprising:
contacting a sample comprising the target protein with a plurality of type 1 magnetic beads capable of recognizing and binding the affinity tag under conditions sufficient for the plurality of type 1 magnetic beads to bind to the affinity tag fused to the target protein in the sample, wherein a protease cleavage sequence is between the affinity tag and the target protein;
washing the plurality of type 1 magnetic beads to remove contaminating material from the sample;
contacting the sample with a cleavage solution containing an endoprotease that cleaves the cleavage sequence to release the target protein from the plurality of type 1 magnetic beads;
contacting the sample with a plurality of type 2 magnetic beads capable of recognizing and binding to the endoprotease under conditions sufficient to bind the endoprotease in the sample, wherein the type 2 magnetic beads do not bind the affinity tag fused to the target protein; and
removing the plurality of type 1 magnetic beads and the plurality of type 2 magnetic beads, thereby purifying the target protein.