Synthesis and characterization of supermacroporous cryogel with immobilized p‐aminobenzenesulfonamide as affinity ligand for the purification of lactoperoxidase from whey

Abstract

This study proposed the development of a monolithic supermacroporous affinity column for direct capture of lactoperoxidase, a glycoprotein present in milk, whey, and colostrum, with several applications due to its wide antimicrobial activity. A poly(acrylamide)-based cryogel was produced by radical co-polymerization of monomers in frozen aqueous solution and activated with p-aminobenzenesulfonamide as a ligand for specific interaction with the lactoperoxidase. The axial liquid dispersion coefficients at different liquid flow rates were determined by measuring residence time distributions using the tracer pulse-response method. The axial dispersion coefficient was low and the height equivalent to theoretical plate was not dependent on the flow velocity. The adsorptive capacity of affinity cryogel was studied as a function of flow velocity and the best condition was 0.9 cm/min. The response surface methodology was applied to optimize the capture of the enzyme, as a function of pH and salt concentration. Higher purification factor value was found at a salt concentration of 80 mmol/L and pH of 8.0 (p < 0.05). There was no influence of the variables under study on the yield (p > 0.05). The results indicated that affinity cryogel is a promising chromatography support for the use in high-throughput one-step purification of lactoperoxidase from whey.

Cryogel with Modular and Clickable Building Blocks: Toward the Ultimate Ideal Macroporous Medium for Bacterial Separation

“…Composite cryogel has great potential in conquering this adverse factor in bacterial separation due to its well-designed macroporous structure (with pore size ranging from 1 to 100 μm). − The interconnected channel and macroporous structure of cryogel are formed in cryo-gelation reactions where ice crystals perform as porogens forming the specific architecture after melting. − The macroporous structure endows the composite with fast mass transfer, making it possible to provide unhindered passage of cells through the monolith even in biological samples with abundances of interfering substances without complicated pretreatment strategies. ,, However, it is still difficult for bacteria to enter the interior structure of cryogels due to their huge cell size. Accordingly, the ligands used for bacteria recognition binding must be modified on the exposed surface of composite cryogels to enhance the recognizing efficiency. , Still, one of the major challenges of using cryogels for bacterial separation is their relatively lower binding capacities. In previous studies, by incorporating functional polymer chains and/or filler particles, the problem can be resolved to some extent. − Ye and co-workers reported a composite cryogel with boronic acid-integrated polymeric building block for the isolation of bacteria from spiked food samples .…”

“…Accordingly, the ligands used for bacteria recognition binding must be modified on the exposed surface of composite cryogels to enhance the recognizing efficiency. 33,34 Still, one of the major challenges of using cryogels for bacterial separation is their relatively lower binding capacities. In previous studies, by incorporating functional polymer chains and/or filler particles, the problem can be resolved to some extent.…”