Performance of phospho-L-tyrosine immobilized onto alginate/polyacrylamide-based cryogels: Effect of ligand coupling on human IgG adsorption and Fab fragments separation

Marcuz, Camila; Mourão, Cecília Alves; Haupt, Karsten; Bueno, Sônia Maria Alves

doi:10.1016/j.jchromb.2021.122530

Cited by 15 publications

(14 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Here, 48 mg/g catalase onto the p(HEMA-co-AGE) cryogels were immobilized without any additional impurities such as Cibacron Blue F3GA and Fe 3+ ions. The monolith cryogels most used as a stationary phase in affinity chromatography were prepared using copolymers of poly-acrylamide (PAAm), glycidyl methacrylate (GMA), and ethylene glycol dimethacrylate (EDMA) [ 41 ]. The PAAm-Alg-AGE was synthesized by the copolymerization of AGE, acrylamide, and alginate with N, N’-Methylenebisacrylamide, and Ca(II) as the crosslinking agents, and the AGE has a conjugated epoxy group that provided the immobilization of P-Tyr.…”

Section: Discussionmentioning

confidence: 99%

Synthesis and Characterization of a New Cryogel Matrix for Covalent Immobilization of Catalase

Altunbaş

Aslan

Kuşat

et al. 2022

Gels

View full text Add to dashboard Cite

The advantages of cryogels for enzyme immobilization applications include their mechanical and chemical robustness, ease of production, superior porosity, and low cost. Currently, many researchers are exploring porous material-based systems for enzyme immobilization that are more efficient and economically viable. Here, poly(2-Hydroxyethyl methacrylate-co-allyl glycidyl ether) (p(HEMA-co-AGE)) cryogel matrices were synthesized via the free radical cryopolymerization method to be employed as the support material. For the immobilization of the catalase enzyme onto the p(HEMA-co-AGE) cryogel matrix (catalase@p(HEMA-co-AGE), the best possible reaction conditions were determined by altering parameters such as pH, catalase initial concentration, and flow rate. The maximum catalase immobilization amount onto the p(HEMA-co-AGE) cryogel was found to be 48 mg/g cryogel. To determine the advantages of the cryogel matrix, e.g., the stability and reusability of the cryogel matrix, the adsorption–desorption cycles for the catalase enzyme were repeated five times using the same cryogel matrix. At the end of the reusability tests, it was found that the cryogel was very stable and maintained its adsorption capacity with the recovery ratio of 93.8 ± 1.2%. Therefore, the p(HEMA-co-AGE) cryogel matrix affords repeated useability, e.g., up to five times, without decreasing its catalase binding capacities significantly and has promising potential for many industrial applications. Cryogels offer clear distinctive advantages over common materials, e.g., micro/nano particles, hydrogels, films, and composites for these applications. At present, many researchers are working on the design of more effective and economically feasible, porous material-based systems for enzyme immobilization

show abstract

Section: Discussionmentioning

confidence: 99%

Synthesis and Characterization of a New Cryogel Matrix for Covalent Immobilization of Catalase

Altunbaş

Aslan

Kuşat

et al. 2022

Gels

View full text Add to dashboard Cite

show abstract

“…Cryogels are polymeric materials synthesized by the polymerization of monomers under freezing conditions [13–14]. It presents epoxy groups on their surfaces by the addition of allyl glycidyl ether that are potential options for the immobilization of ligand groups due to their high reactivity and formation of covalent bonds with amine, thiol, or hydroxyl groups [15].…”

Section: Introductionmentioning

confidence: 99%

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

Maciel

Mól

Veríssimo

et al. 2022

J of Separation Science

View full text Add to dashboard Cite

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.

show abstract

“…Furthermore, the addition of certain chemicals during cryogel preparation could provide reactive groups for cryogels. In the case of polyacrylamide cryogels, for example, allyl glycidyl ether (AGE) is often used as a functional copolymer monomer for epoxy groups and as the basis for ligand immobilization 25,26 . These advantages and applications made cryogels to be used as a material for immobilizing ligands in biopanning.…”

Section: Introductionmentioning

confidence: 99%

The specific biopanning of single‐domain antibody against haptens based on a functionalized cryogel

Zhang

Liu

Han

et al. 2022

J of Molecular Recognition

View full text Add to dashboard Cite

Phage display technology is commonly applied for high-throughput screening of single-domain antibodies (sdAbs), and the problem of non-specific adsorption caused by carrier proteins seriously affects the biopanning of single-domain antibodies specific to haptens. In this paper, enrofloxacin (ENR)-functionalized cryogels were prepared by the ethylenediamine (EDA) and carbodiimide methods for application in the biopanning of ENR-specific phages. To improve the efficiency of biopanning, double blocking, a wash solution flow rate of 1 mL/min, and phage pre-incubation were applied to the biopanning process through single-factor experiments. Results of flat colony counting showed that the phage output of AG-ENR cryogels was 15 times higher than that of AG cryogels for the same input amount. And seven complete sequences of ENR-specific shark sdAbs were obtained by monoclonal phage ELISA and sequence alignment. All these results indicate that functionalized cryogels could be used as a novel and efficient method for phage biopanning for single-domain antibodies to haptens.

show abstract

Performance of phospho-L-tyrosine immobilized onto alginate/polyacrylamide-based cryogels: Effect of ligand coupling on human IgG adsorption and Fab fragments separation

Cited by 15 publications

References 40 publications

Synthesis and Characterization of a New Cryogel Matrix for Covalent Immobilization of Catalase

Synthesis and Characterization of a New Cryogel Matrix for Covalent Immobilization of Catalase

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

The specific biopanning of single‐domain antibody against haptens based on a functionalized cryogel

Contact Info

Product

Resources

About