Designing a Multifunctional Chromatography Stationary Phase for (Bio)Product Purification via Green Chemistry
(2024) KBKM01 20241Pure and Applied Biochemistry
- Abstract
- The multifunctional cryogels were synthesized by atom transfer radical polymerization (ATRP) using hemoglobin as a biocatalyst (ATRPase) and post-modified to purify hemoglobin and glycoprotein simultaneously. Meanwhile, the optimization of purified glycoprotein was also conducted by introducing epoxy group nanoparticles. The nanoparticle concentrated concanavalin A, the ligand, to adsorb glycoprotein effectively. Scanning electron microscopy (SEM) was used to characterize the morphology of cryogels with/without nanoparticles. Batch adsorption and kinetics adsorption were used to evaluate the adsorption capacity and behavior of the chromatography columns. The Langmuir and bis-Langmuir models were used to fit the batch adsorption of... (More)
- The multifunctional cryogels were synthesized by atom transfer radical polymerization (ATRP) using hemoglobin as a biocatalyst (ATRPase) and post-modified to purify hemoglobin and glycoprotein simultaneously. Meanwhile, the optimization of purified glycoprotein was also conducted by introducing epoxy group nanoparticles. The nanoparticle concentrated concanavalin A, the ligand, to adsorb glycoprotein effectively. Scanning electron microscopy (SEM) was used to characterize the morphology of cryogels with/without nanoparticles. Batch adsorption and kinetics adsorption were used to evaluate the adsorption capacity and behavior of the chromatography columns. The Langmuir and bis-Langmuir models were used to fit the batch adsorption of glycoprotein and hemoglobin, respectively. For kinetics adsorption, both proteins were followed by the pseudo-second-order model. Based on these models, the theoretical maximum capacity was calculated and compared. The maximum capacity of hemoglobin was 148.58 and 309.70 mg protein/mL cryogels for cryogels with/without nanoparticles; The maximum capacity of ovalbumin, a glycoprotein, was 93.98 and 43.63 mg protein/mL cryogels for cryogels with/without nanoparticles. It was found that introducing nanoparticles can enhance the efficiency of glycoprotein purification by increasing the concentration of immobilized ligands, which increased 50.35 mg protein/mL cryogels. However, immobilizing nanoparticles impacted hemoglobin adsorption and reduced it. (Less)
- Popular Abstract
- Biotechnology is becoming increasingly important in our society, enabling the production of many valuable substances across various fields. Microorganisms like cells, yeast, and bacteria are used to create bioproducts for medicine, food production, material development, and more. However, these bioproducts often contain impurities, making it essential to obtain high-purity products. This is done through a process called downstream processing.
One key technique in downstream processing is chromatography. Chromatography effectively separates impurities from target products and has two phases: the mobile phase (a solution) and the stationary phase (column). As the mobile phase moves through the column, wanted molecules are captured, allowing... (More) - Biotechnology is becoming increasingly important in our society, enabling the production of many valuable substances across various fields. Microorganisms like cells, yeast, and bacteria are used to create bioproducts for medicine, food production, material development, and more. However, these bioproducts often contain impurities, making it essential to obtain high-purity products. This is done through a process called downstream processing.
One key technique in downstream processing is chromatography. Chromatography effectively separates impurities from target products and has two phases: the mobile phase (a solution) and the stationary phase (column). As the mobile phase moves through the column, wanted molecules are captured, allowing for efficient purification. By optimizing this process, we can obtain high-purity bioproducts more efficiently.
In this project, we designed the stationary phase to capture multiple proteins simultaneously, using a more sustainable approach to create the chromatography column. We chose cryogels because their unique macroporous structure allows large molecules to pass through directly, reducing the need for sample preparation and shortening the process time. We developed a green method to synthesize these cryogels, using hemoglobin as a catalyst instead of metals, a novel approach that reduces environmental impact. We also generated multiple binding sites within the cryogels using various techniques. The target proteins in this study were hemoglobin and a glycoprotein called ovalbumin. We further optimized the system by introducing nanoparticles, which provided more binding sites for capturing glycoproteins.
The multifunctional cryogels developed in this project offer a more efficient and sustainable method for protein purification, demonstrating enhanced effectiveness in capturing target proteins. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9167848
- author
- Lei, Xiaolin LU
- supervisor
- organization
- course
- KBKM01 20241
- year
- 2024
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- biochemistry, cryogels, protein purification, chromatography, green chemistry
- language
- English
- id
- 9167848
- date added to LUP
- 2024-07-03 09:35:06
- date last changed
- 2024-07-03 09:35:06
@misc{9167848,
abstract = {{The multifunctional cryogels were synthesized by atom transfer radical polymerization (ATRP) using hemoglobin as a biocatalyst (ATRPase) and post-modified to purify hemoglobin and glycoprotein simultaneously. Meanwhile, the optimization of purified glycoprotein was also conducted by introducing epoxy group nanoparticles. The nanoparticle concentrated concanavalin A, the ligand, to adsorb glycoprotein effectively. Scanning electron microscopy (SEM) was used to characterize the morphology of cryogels with/without nanoparticles. Batch adsorption and kinetics adsorption were used to evaluate the adsorption capacity and behavior of the chromatography columns. The Langmuir and bis-Langmuir models were used to fit the batch adsorption of glycoprotein and hemoglobin, respectively. For kinetics adsorption, both proteins were followed by the pseudo-second-order model. Based on these models, the theoretical maximum capacity was calculated and compared. The maximum capacity of hemoglobin was 148.58 and 309.70 mg protein/mL cryogels for cryogels with/without nanoparticles; The maximum capacity of ovalbumin, a glycoprotein, was 93.98 and 43.63 mg protein/mL cryogels for cryogels with/without nanoparticles. It was found that introducing nanoparticles can enhance the efficiency of glycoprotein purification by increasing the concentration of immobilized ligands, which increased 50.35 mg protein/mL cryogels. However, immobilizing nanoparticles impacted hemoglobin adsorption and reduced it.}},
author = {{Lei, Xiaolin}},
language = {{eng}},
note = {{Student Paper}},
title = {{Designing a Multifunctional Chromatography Stationary Phase for (Bio)Product Purification via Green Chemistry}},
year = {{2024}},
}