Practical Protein Chemistry - A. Darbre 1989

Affinity Chromatography of Proteins
Methods for Preparation of Affinity Sorbent
Concept of Affinity Sorbent

Selecting a sorbent with appropriate characteristics is of paramount importance in Affinity Chromatography. An affinity sorbent consists of three components: a matrix, a spacer arm, and a Ligand. The presence of all three elements is not always strictly necessary; for instance, in the affinity chromatography of Lectins, the matrix itself (Sephadex G-75, containing glucose and mannose residues) serves as an effective ligand recognized by the lectin. Another example is the purification of anti-dextran Antibodies on Sephadex G-100. Typically, however, the ligand is immobilized on the matrix via an intermediate spacer arm, the function of which is to project the ligand away from the sorbent surface, thereby eliminating steric hindrance during its interaction with the target protein.

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FIG. 5.2. Various Applications of Affinity Chromatography within a general Protein Purification scheme.



Last update: 06/08/2026

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