Cyanogen bromide agarose, activation

March, S.C., Parikh, I., and Cuatrecasas, P. (1974) A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal. Biochem. 60, 149-152. 

It should be mentioned here that another family of hydrocarbonaceous phases has been developed for the hydrophobic chromatography of biopolymers 49, 50). For the preparation of such stationary phases agarose beads are first activated with cyanogen bromide and subsequently reacted... 

Coupling of Protein to Cyanogen Bromide-Activated Agarose... 

If purchased, cyanogen bromide-activated agarose will be in a dried form in the presence of stabilizers and will require swelling and washing before use. The activated groups are liable to hydrolysis at alkaline pH and thus the swelling and washing is carried out at acid pH. 

The most commonly used biopolymers, such as agarose, contain alcoholic hydroxyl groups which can be activated with cyanogen bromide however, better methods have recently been developed including activation with sulfonyl chlorides (17), 2-fluoro-l-methylpyridinium toluene sulfonate (FMP) (10), and chlorocarbonates (18). The first two are commercially available as activated supports tresyl-activated Sepharose (Pharmacia) and FMP-Trisacryl (BioProbe International). The newer methods yield more stable bonds, which preclude leaching of the enzyme from the matrix. Most of these activated supports are too expensive for commercial use in a large process bioreactor however, they may be extremely useful for preparing analytical bioreactors. 

A chromatographic matrix with free OH groups, such as agarose, is first activated with cyanogen bromide (CNBr). Ligand is then coupled to the... 

Figure 7-5. Effect of pH on the amount of alanine covalently bound to cyanogen bromide activated agarose. [From P. Cu-atrecasas, J. Biol. Chem., 245 3059 (1970).]...

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