Immobilization of Affinity Ligands

Affinity ligands are immobilized through covalent binding to the support material. A dense, stable coverage of the support is desired, and most methods for immobilization use two steps support activation and coupling to the affinity ligand (see Section 4.2 regarding enzyme immobilization methods). 

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Salt-induced immobilization of affinity ligands onto epoxide-activated supports,... 

Wiley, J.P., Hughes, K.A., Kaiser, R.J., Kesicki, E.A., Lund, K.P., and Stolowitz, M.L. (2001) Phenylboronic acid-salicylhydroxamic acid bioconjugates. 2. Polyvalent immobilization of protein ligands for affinity chromatography. Bioconjugate Chem. 12, 240-250. 

Table 3.19. Some forms of affinity chromatography that may be employed to purify a selected protein. Virtually all proteins carry out their biological effects by interacting in some way with other molecules, which can be given the general title ligands . This interaction is often quite biospecific. Immobilization of such ligands (or molecules which mimic ligands) onto chromatographic beads thus facilitates selective purification of the molecule of interest...

Several techniques were developed for immobilization of the ligands in affinity gels. 

Affinity chromatography involves the immobilization of a ligand (substrate, analog, inhibitor or effector) on agarose as a support that can be used selectively to adsorb the enzyme from a sample solution passed over it. 

Fig. 2 Common immobilization effects during the coupling of affinity ligands to solid supports.

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