Affinity purification methods involving imidazole elution

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Separation or purification

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530412, 5303871, 5303911, 5483351, C07K 1322, C07K 1600, C07K 1646, C07D23354

Patent

active

057262934

ABSTRACT:
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 affinity complex formation; (iii) isolating the complex; (iv) treating the complex with imidazole to disrupt the complex, causing the release of the protein from the complex; and (v) isolating the protein. According to one embodiment, 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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