Integrin 51: A new purification strategy based on immobilized peptides
Novel efficient and robust affinity chromatography material: There are several strategies known for the purification of integrins by affinity chromatography, but the disadvantages of common strategies like insufficient selectivity or compelling conditions for the elution still require alternatives. A new strategy, based on the immobilized C-terminally modified peptide Ac-Gly-Ala-c-(CysSS-Arg-Arg-Glu-Thr-Ala-Trp-Ala-Cys SS)-Gly-Ala-O(CH2CH2O)2CH 2CH2-NH2 allows for the affinity purification of the integrin 51. While RGD peptides have been proven in the past to be inappropriate for selective purification of integrins by affinity chromatography, the new peptide can be efficiently used for selective enrichment of the integrin 51. It is a specific ligand of the target protein, but does not contain an RGD sequence. The application of well-characterized affinity chromatography material with a site-specifically immobilized peptide allows to obtain integrin 51 in a single chroma tography step without contamination by other integrins. This process combines the advantages of a selective and monospecific protein-ligand recognition with mild elution conditions and a low sensitivity of the immobilized ligand with respect to column regeneration.