Affinity Chromatography - Columns

Affinity Chromatography Columns

Affinity chromatography columns separate proteins by providing a stationary phase modified with a ligand that specifically interacts with the protein.

Binding: The target molecule is specifically and reversibly adsorbed by a complementary ligand and immobilized on a matrix. Examples of a complementary ligand include an inhibitor, substrate analog or cofactor, or an antibody which specifically recognizes the target molecule. The selectivity is often based on spatial recognition, a ‘lock-and-key’ mechanism.

Elution: The adsorbed molecule is eluted either by competitive displacement or a conformation change through a shift in pH or ionic strength. Typical molecular pairs are antigens and antibodies, enzymes and coenzymes, and sugars with lectins.

TSKgel analytical affinity columns and their applications

TSKgel Protein A-5PW for mAb titer determination

The column uses a Protein A ligand which specifically binds the Fc part of antibodies. Therefore, antibodies are captured while contaminants flow through. Mab concentration is then determined by eluting and detecting it.

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TSKgel FcR-IIIA-affinity columns for Fc receptor affinity studies of antibodies

The columns employ a recombinant version of the Fc gamma IIIa receptor. They separate monoclonal antibodies according their affinity to the receptor and enable a fast analysis of biological activity as well as fractionation along with biological activity.

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TSKgel affinity columns for modification with ligands of choice

The choice of a specific ligand is dictated by the expected interaction between the sample and the bonded phase of the resin used. The TSKgel affinity column line consists of three group specific ligands and one chemically active functionality.

Well known applications for each type of TSKgel affinity column:

Group specific ligands Application
TSKgel Boronate-5PW carbohydrates, nucleic acids, nucleosides, nucleotides, catecholamines
TSKgel Chelate-5PW immunoglobulins, transferrin, lectins, milk proteins, membrane proteins, peptides


Chemically activated ligands Application
TSKgel Tresyl-5PW coupling of ligands to form a custom affinity resin 

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