Affinity purification methods involving amino acid mimetics...

C - Chemistry – Metallurgy – 07 – K

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C07K 19/00 (2006.01) C07K 1/14 (2006.01) C07K 1/22 (2006.01) C07K 16/00 (2006.01) C07K 16/06 (2006.01)

Patent

CA 2146162

Disclosed is a method of isolating a protein from a sample, involving (i) providing a first molecule which is capable of forming an affinity complex with the protein; (ii) contacting the sample with the first molecule under conditions which allow af- finity complex formation; (iii) isolating the complex; (iv) treating the complex with a second molecule, the second molecule mi- micking an amino acid residue of either the protein or the first molecule which is critical to the complex formation, so that the second molecule disrupts the complex; causing the release of the protein from the complex; and (v) isolating the protein. Accord- ing to one embodiment, the amino acid mimetic imidazole is used as a very gentle elution reagent to disrupt a protein A-antibody fusion protein complex, a technique which has general application for the isolation of antibodies or recombinant antibody fusion proteins.

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