Double-layer vs single-layer cryogels as a dye affinity chromatography column, comparison study
(2022) KEMR30 20221Department of Chemistry
- Abstract
- Dye affinity chromatography is one of the chromatography techniques which uses dye as a ligand for protein purification. In this study, single-layer poly(acrylamide-allyl glycidyl ether) cryogel was prepared, which contains free epoxy groups at sub-zero temperature. The double-layer was prepared inside a primary single-layer cryogel to form a secondary network inside the gel. Cibacron Blue F3GA was immobilized onto single and double-layer cryogels by covalently binding with the epoxy groups. Bovine serum albumin (BSA) adsorption on the formed dye affinity single and double-layer cryogels was investigated. It was found that the kinetic model and adsorption isotherms follow the Pseudo-second-order and Freundlich model, respectively. The data... (More)
- Dye affinity chromatography is one of the chromatography techniques which uses dye as a ligand for protein purification. In this study, single-layer poly(acrylamide-allyl glycidyl ether) cryogel was prepared, which contains free epoxy groups at sub-zero temperature. The double-layer was prepared inside a primary single-layer cryogel to form a secondary network inside the gel. Cibacron Blue F3GA was immobilized onto single and double-layer cryogels by covalently binding with the epoxy groups. Bovine serum albumin (BSA) adsorption on the formed dye affinity single and double-layer cryogels was investigated. It was found that the kinetic model and adsorption isotherms follow the Pseudo-second-order and Freundlich model, respectively. The data showed that the binding capacity of double-layer cryogel decreased compared to single-layer cryogel due to lower surface area. However, the mechanical stability of the double-layer cryogel (based on Young's modulus) is higher than that of the corresponding single-layer cryogel. Both single and double-layer cryogels could be reused more than ten times in adsorption/elution cycles without losing their binding capacity. The physical and chemical properties of the cryogels were characterized by scanning electron microscopy, confocal microscopy, krypton adsorption, fourier transform infrared (FT-IR) spectroscopy, and texture analysis. (Less)
- Popular Abstract
- Seperation and purification of the biological molecules from the complex mixture of many substances is a keystone phase of modern process biotechnology. The cost of current bioseparation methods is high in large scale manufacturing and the demand to obtain highly purified biomolecules like proteins is more and more increasing. These two critical points stimulate scientists to research and develop new polymeric materials to be used in bioseperation.
Supermacroporous cryogels have drawn significant attention in the bioseparation applications due to the porous structure, efficient permeability, and the possibility to form them from the biodegradable materials. Cryogels are synthesized by freezing polymerization techniques at temperature of... (More) - Seperation and purification of the biological molecules from the complex mixture of many substances is a keystone phase of modern process biotechnology. The cost of current bioseparation methods is high in large scale manufacturing and the demand to obtain highly purified biomolecules like proteins is more and more increasing. These two critical points stimulate scientists to research and develop new polymeric materials to be used in bioseperation.
Supermacroporous cryogels have drawn significant attention in the bioseparation applications due to the porous structure, efficient permeability, and the possibility to form them from the biodegradable materials. Cryogels are synthesized by freezing polymerization techniques at temperature of -12 ºC. The solution containing monomers, cross-linkers, initiators is frozen and then consists of the ice crystals and non-frozen liquid between the ice crystals. The polymerization reaction occurs in this non-frozen liquid and the ice crystals grow during freezing. After thawing of the ice crystals, permanent macroporous structures with interconnected channels is formed.
Cryogel column is widely used in affinity chromatography for purification. However, the efficient mass transfer and high flow rate properties of cryogels are the reasons for the reduction of surface area of the network, thefore decreasing the binding capacity for small-sized molecules. One of other biggest limitations of cryogels is their mechanical weakness due to their low-density structure.
With the aim to further improve the surface area and mechanical stability of cryogels, double-layer cryogels were synthesized inside the primary network of cryogels using the cryogelation method. The secondary layer of cryogels was absorbed in the pore walls and was formed inside and on the surface of the primary walls.
In this work, the comparison of binding capacity towards bovine serum albumin (BSA) and mechanical stability between single-layer and double-layer cryogels was studied. The results show that the mechanical stability of double-layer cryogels increased up to 7-compared to the single-layer cryogels. However, the binding capacity of double-layer cryogel is lower than that of single-layer cryogel due to lower surface area. Both single and double-layer cryogels could be reused more than ten times in adsorption/elution cycles without losing their binding capacity.
In conclusion, although cryogels still have some limitations in biotechnology applications, which are neccessary paid atention in the future work, they are attractive for scientist in many areas of chemistry, biology, biotechnology and bioengineering due to their cheap materials and reusibility. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9079984
- author
- Trinh, Thi Hoai Thu LU
- supervisor
- organization
- course
- KEMR30 20221
- year
- 2022
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- dye-affinity chromatography, single-layer cryogels, double-layer cryogels, protein purification, bovine serum albumin, biochemistry
- language
- English
- id
- 9079984
- date added to LUP
- 2022-06-02 14:34:04
- date last changed
- 2022-06-02 14:34:04
@misc{9079984,
abstract = {{Dye affinity chromatography is one of the chromatography techniques which uses dye as a ligand for protein purification. In this study, single-layer poly(acrylamide-allyl glycidyl ether) cryogel was prepared, which contains free epoxy groups at sub-zero temperature. The double-layer was prepared inside a primary single-layer cryogel to form a secondary network inside the gel. Cibacron Blue F3GA was immobilized onto single and double-layer cryogels by covalently binding with the epoxy groups. Bovine serum albumin (BSA) adsorption on the formed dye affinity single and double-layer cryogels was investigated. It was found that the kinetic model and adsorption isotherms follow the Pseudo-second-order and Freundlich model, respectively. The data showed that the binding capacity of double-layer cryogel decreased compared to single-layer cryogel due to lower surface area. However, the mechanical stability of the double-layer cryogel (based on Young's modulus) is higher than that of the corresponding single-layer cryogel. Both single and double-layer cryogels could be reused more than ten times in adsorption/elution cycles without losing their binding capacity. The physical and chemical properties of the cryogels were characterized by scanning electron microscopy, confocal microscopy, krypton adsorption, fourier transform infrared (FT-IR) spectroscopy, and texture analysis.}},
author = {{Trinh, Thi Hoai Thu}},
language = {{eng}},
note = {{Student Paper}},
title = {{Double-layer vs single-layer cryogels as a dye affinity chromatography column, comparison study}},
year = {{2022}},
}