Enhancements in sugar immobilization in polymeric macroporous matrices for affinity capture

Abstract

The use of macroporous monolithic matrices in the purification of biocompounds is constantly growing and developing. In this work, the objective was to optimize the quantity of N-acetyl-D-glucosamine (D-GlcNAc) immobilized on the surface of macroporous polymeric cryogels for capture of lectins from less clarified solutions. Surface response methodology was applied and it was observed that the immobilization temperature of the glutaraldehyde (GLU) and the D-GlcNAc concentration influenced the amount of sugar immobilized. The matrices produced with 1.1% of allyl glycidyl ether were functionalized by GLU. Optimal maximum condition was obtained with mean value of 160.39 AE 26.38 mg of D-GlcNAc immobilized per gram of dry cryogel. Characterization analyses of the matrices showed that the activation process was effective, maintaining the macroporous structure and physical characteristics. The adsorbents produced were tested for capture of lectins from a crude protein solution of barley. At tested conditions, adsorbent capture around 11% of protein in solution but reduce the hemaglutinating capacity in 40%, demonstrating its selectivity. The cryogels functionalized with D-GlcNAc present potential for use in capture compounds by affinity with carbohydrates, such as lectins.

Lectin Purification through Affinity Chromatography Exploiting Macroporous Monolithic Adsorbents

“…On the other hand, these same ions at high concentrations are capable of agglutinating cells, as observed by Silva et al [15] in their study on the influence of Ca 2+ and Mn 2+ ions for their hemagglutinating activity of cassava leaf lectin. Thus, to ensure that the agglutination is being mediated by lectin and not by excess ions, it is necessary to inhibit this activity with the addition of carbohydrates [15].…”

“…Marine sponge [47] Ion exchange chromatography Jacalina (Artocarpus integrifólia) Jackfruit [15] Affinity chromatography Con A Pork beans [26] Gel filtration chromatography PeCL Fungus (Penicillium chrysogenum) [41] Affinity chromatography followed by ion exchange…”

“…The principle of the method in question is to improve the separation capacity of biomolecules from specific, non-covalent bonds to insoluble supports, favoring the obtaining of bio-separations with high selectivity [12][13][14]. It is known that the use of monolithic, macroporous matrices applied in the purification of bio compounds is in constant growth and development [15]. In this sense, this study aimed to report and discuss information from the literature on polymeric, macroporous, monolithic adsorbents applied in the purification of lectins using affinity chromatography.…”