Affinity Gels

Chromatography

Affinity gel chromatography which aim to purify molecule by non-covalent and reversible binding is in the most cases the method of choice to obtain desired lectins or conversely oligo-polysaccharides or glycoconjugates with high yield and purity. These reagents are provided either as bulk or prepacked into 1 to 5 mL columns.

Two kind of affinity gel are proposed by GLYcoDiag with different specificities :
Carbohydrate affinity gel (CarbPROFILE gel) or Lectin affinity gel (LEctPROFILE gel).

Carbohydrate affinity gel (CarbPROFILE gel)

CarbPROFILE gels are monosaccharides-Sepharose affinity matrices used for purification of specific carbohydrate-binding proteins. The carbohydrates are attached through their non reducing hydroxyl group after pre-activation of sepharose matrix by divinylsulfone (DVS) (see scheme 1 below). The binding of lectins and carbohydrates binding proteins to carbohydrate affinity gel is non-covalent and reversible with high capacity. Lectins and carbohydrates binding proteins are both usually stable compounds which can be recovered by competitive elution (i.e. 0.2 to 0.5 M of monosaccharide) or by modulations of pH and/or ionic strength in high yield and purity.

Scheme 1. CarbPROFILE gel matrix.

REFERENCE NAME SPECIFICITY CAPACITY UNIT SIZE a (mL)
ManGel Mannose-CarbPROFILE gel Mannose binding protein >30 (based on ConA lectin) 5, 10 or 25
GalGel Galactose-CarbPROFILE gel Galactose binding protein >15 (based on AIA lectin) 5, 10 or 25
GlcNAcGel N-Acetylglucosamine-CarbPROFILE gel N-Acetylglucosamine binding protein >15 (based on WGA lectin) 5, 10 or 25

Table 1. Specifications of CarbPROFILE gel.  a. available in a prepacked column or in suspension.

Applications

  • Purification of large range of glycoproteins which recognize specific carbohydrates moeities.

Lectin affinity gel (LEctPROFILE gel)

LEctPROFILE gels are affinity gel chromatography where lectins are immobilized on a Sepharose 4B fast flow matrix. Glycoconjugates can be recovered by competition with the specific inhibitory monosaccharide of the lectin. LEctPROFILE gels are used for the purification of glycoconjugates with specific N-glycan residues. LEctPROFILE gels are personnalized reagents produced on your request (see our list of available lectins).

Scheme 2. LEctPROFILE gel matrix

NAME SPECIFICITY BINDING CAPACITY UNIT SIZE b (mL)
LEctPROFILE gel a ≥1 1, 2 or 5

Table 2. Specifications of LEctPROFILE gel.  a. available in a pre-packed column or in suspension;  b. For specificity of LEctPROFILE gel, see our lectins  specificities table.

Applications

  • Purification of polysaccharides.
  • Purification of complex glycoproteins having specific glycans structures.