Affinity Columns

TSKgel Affinity Chromatography Columns

The coupling of m-aminophenyl boronate to a polymeric support results in the TSKgel Boronate-5PW column. The coupling makes a ligand capable of forming a tetrahedral boronate anion under alkaline pH conditions that can bind with 1,2 cis-diol groups such as those found in carbohydrates, carbohydrate-containing compounds and catecholamines. Adsorption onto the TSKgel Boronate-5PW matrix takes place in basic buffers such as HEPES and morpholine while desorption takes place in carbohydrate or amine-containing mobile phases like sorbitol or Tris.

TSKgel Chelate-5PW columns contain iminodiacetic acid (IDA) groups that are covalently bonded to a polymeric support. Prior to chromatography, a metal ion, such as Zn²⁺, Ni²⁺ or Cu²⁺, is chelated to the IDA group. The TSKgel Chelate-5PW column has high binding capacity for the target molecules. Peptides and proteins containing histidine residues will normally adsorb to these chelated ions at neutral pH. A buffer containing imidazole or glycine is used for protein desorption. The key to making successful use of this retention mechanism is selecting the proper metal ion and elution buffer.

Unlike other TSKgel affinity columns, the TSKgel Tresyl-5PW columns which are derivatized with the 2,2,2-trifluroethanesulfonyl ligand, require activation with a user-selected ligand containing amino, thiol, phenol, or imidazole groups. The resulting structure is literally a custom affinity ligand with excellent pH stability and minimal ligand loss due to leaching. TSKgel Tresyl-5PW readily reacts with amino or thiol groups to form stable covalent alkylamines or thioethers.