Tresyl activation

Figure 14.14 Additional hydroxyl-particle activation methods include bis-epoxide modification, tosyl activation, and tresyl activation methods. The tosyl chloride and tresyl chloride activation procedures must be done in dry organic solvent, but the coupling of an amine-containing ligand can be done in either organic solvent or aqueous buffer.

Soluble enzymes, unless stated otherwise. Enzyme immobilized on tresyl-activated Sepharose. 

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. 

Tresyl activation results in a connection to a protein via a secondary amine bond. 

The advantage of this factorial design technique is the ability to examine the influence of aU immobilization parameters independently as they are orthogonal. Figure 14 indicates that half of the maximum binding capacity is achieved after 30 min, which means that the Tresyl activation is a fast reaction. 

Tresyl activation Carbodiimide activation Glutaraldehyde Epoxide... 

McConway MG and Chapman RS. Application of solid-phase antibodies to radioimmunoassay evaluation of two pol3mieric microparticles, dynospheres and nylon, activated by carbonyldiimidazole or tresyl chloride. J. Immunol. Methods 1986 95 259-266. 

A simple method for the activation of hydroxyl group carrying supports such as agarose, cellulose, diol-silica, glycophase-glass or hydroxyalkyl methacrylate copolymer with 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride) was introduced by Nilsson and Mosbach [108] ... 

Figure 14-1 Functionalization and activation of electrode materials, a) silanization of supports bearing hydroxylic groups b) reaction with cyanuric chloride c) activation with cyanogen bromide d) binding via amide bonds after activation with 1,1-carbonyldiimidazole e) activation with good leaving groups such as tresyl chloride f) formation of Schiff bases between aldehyde groups on the support and amino groups at the biomolecule g) activation of carboxylic groups with carbodiimides or h) via the acid chlorides.

Figure 4.5-1. Chemical structures of activated PEGs for amine conjugation (a) PEG-dichlorotriazine (b) PEG-tresylate (c) PEG-aldehydes (d) PEG-succinimidyl succinate (e) PEG-imidazole carbonate (f) PEG-phenyl carbonate (g) PEG-succinimidyl carbonate (h) PEG-benzotriazole carbonate and (i) PEG-active ester.

Other activation methods are performed with carbonyldiimidazole or tresyl chloride (14) both jdeld uncharged materials. Coupling with carbonyldiimidazole (CDI) is very rapid, but the bond formed, a substituted carbamate, is not as stable as the secondary amine formed with the slower tresyl chloride. 

For initial activation with CNBr, tresyl chloride etc. Proteins, peptides... 

It is advisable to have some ready-to-use commercially available affinity columns to hand, such as IMAC resins, p-aminobenzamidine. Reactive Blue, heparin. Protein A for antibody purification, as well as some activated resins such as tresyl, epoxy or N-hydroxysuccinimide. 

K. Nilsson, K. Mosbach, Tresyl chloride-activated supports for enzyme immobilization, Methods Enzymol. 135 (1987), 65-78. 

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