Patent ID: 6723510
Filing Date: 2004-04-20
Classification: C07K

Abstract:
A method for isolating a fusion protein, wherein said fusion protein comprises a peptide, polypeptide, or protein and an affinity peptide tag consisting of a plurality of consecutive histidine residues from a sample in a vessel, comprising the steps of:(a) combining the sample containing the fusion protein with metal-chelate affinity particles suitable for binding said fusion protein, said metal-chelate affinity particles being insoluble in the sample; (b) collecting the metal-chelate affinity particles; (c) separating the metal-chelate affinity particles from the unbound remainder of the sample; (d) optionally, resuspending the metal-chelate affinity particles in a solution; (e) optionally, eluting said fusion protein from the metal-chelate affinity particles, followed by separating the metal-chelate affinity particles from said eluted fusion protein; wherein at least one of steps (a), (b), (c), (d) if present, and (e) if present is performed in the presence of 0.0005%-2% (v/v) detergent sufficient to reduce loss of metal-chelate affinity particles during any separation or collection step, in comparison to the same method performed in the absence of detergent.