Patent ID: 6326155
Filing Date: 2001-12-04
Classification: C40B,G01N

Abstract:
A method for isolating an affinity ligand suitable for separating a target molecule from a solution containing said target molecule, wherein said affinity ligand is a polypeptide, the method comprising:(a) selecting, with respect to the target molecule, a first solution condition at which an affinity ligand will bind to said target molecule and at which the target molecule is not denatured;(b) selecting, with respect to the target molecule, a second solution condition at which the affinity ligand will not bind to said target molecule and at which the target molecule is not denatured, wherein said second solution condition is different from said first solution condition;(c) providing a lebrary of polypeptides;(d) contacting said library of polypeptides with said target molecule at the first solution condition, for sufficient time to permit polypeptide/target binding complexes to form;(e) removing polypeptides that do not bind the target under the first solution condition;(f) altering the first solution condition of contacting step (d) to the second solution condition; and(g) recovering polypeptides that release the target molecule under the second solution condition, wherein the recovered polypeptides identify isolated affinity ligands that bind said target molecule under said first solution condition and do not bind said target molecule under said second solution condition.