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