US EPA

5202255 

Journal Article 

Adsorbents for affinity chromatography. Use of N-hydroxysuccinimide esters of agarose 

Cuatrecasas, P; Parikh, I 

1972 

Yes 

Biochemistry
ISSN: 0006-2960
EISSN: 1520-4995 

11 

12 

2291-2299 

English 

5028497 

10.1021/bi00762a013 

Procedures are described for the preparation of the N-hydroxysuccimmide ester of succinylated aminoalkyl agarose derivatives. These active ester derivatives of agarose are stable for months when stored in dioxane. The reaction of the active ester of agarose with various amino acids and with trypsin has been studied in detail. Stable amide linkages are formed very rapidly (at 4°) with the unprotonated form of primary aliphatic or aromatic amino groups in the pH range of 6-9. Of the amino acid functional groups tested, only sulfhydryl groups compete effectively with free amino groups for reaction. The iV-hydroxysuccinimide ester derivative of agarose can be used by very simple and mild procedures to immobilize proteins and complex amino group containing ligands to agarose. The coupled products are separated from the matrix backbone by lengthy hydrocarbon extensions. Similar procedures can be used to attach ligands and proteins to porous glass beads. These studies in addition demonstrate that agarose beads tolerate quite well certain organic solvents such as dioxane and methanol. The activated agarose derivatives, after lyophilization to remove dioxane, are quite stable in the powder form and readily swell and react with amino groups when suspended in aqueous medium. 1972, American Chemical Society. All rights reserved.