Affinity chromatography using biotinylated chromatographic probes

Affinity chromatography using biotinylated chromatographic probes 

The procedure for coupling 3a to the peptidyl-resin is similar to that for probe 2. After acid cleavage, a solution of the crude mixture is applied directly to an avidin-based affinity column and elution with buffers removes all underivatized chains. The peptide-containing column can be treated with an organic base to release free peptide (Protocol 5) or used directly for binding studies or affinity purification. 

Protocol 5. Purification of peptides using biotinylated-probe 3 

At the end of polypeptide chain assembly, take enough peptidyl-resin so as to have about 3 p-mol protected peptide and remove the N -protection from the terminal residue as usual. Wash the peptidyl-resin with DCM. If chain assembly is performed with Boc chemistry, neutralize 2 min) the freshly deprotected peptidyl-resin with 5% DIPEA in DCM and then wash with DCM. 

Transfer the solution to the reaction vessel containing the N-deprotected peptidyl-resin and vortex at room temperature overnight. 

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Affinity chromatography using biotinylated chromatographic probes... 

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