Design and application of a newly generated bio/synthetic cryogel column for DNA capturing

Önal, Burcu; Odabaşı, Mehmet

doi:10.1007/s00289-020-03387-x

Cited by 8 publications

(5 citation statements)

References 61 publications

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“…After optimization, a binding capacity of 39.7 mg of pDNA/g of cryogel was obtained, as well as an Abs260/280 ratio of 1.85, indicating that the eluted DNA has a high degree of purity. Even though this novel affinity approach provides a high binding capacity, further studies need to be conducted to evaluate its performance (purity and recovery) for sc isolation and impurity (RNA, host proteins, gDNA, endotoxins) removal [ 121 ].…”

Section: Cryogelsmentioning

confidence: 99%

“…Immobilized metallic chelate affinity (IMAC) technology was implemented into cryogel supports for pDNA isolation from a lysate sample by Önal and Odabaşı (2021), where an Fe 3+ , extracellular polymeric substances, and PHEMA composite cryogel were used. After optimization, a binding capacity of 39.7 mg of pDNA/g of cryogel was obtained, as well as an Abs260/280 ratio of 1.85, indicating that the eluted DNA has a high degree of purity.…”

Section: Dna Purificationmentioning

confidence: 99%

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Cryogels and Monoliths: Promising Tools for Chromatographic Purification of Nucleic Acids

Ribeiro,

Luís,

Rodrigues

et al. 2024

Gels

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The increasing demand for highly pure biopharmaceuticals has put significant pressure on the biotechnological industry to innovate in production and purification processes. Nucleic acid purification, crucial for gene therapy and vaccine production, presents challenges due to the unique physical and chemical properties of these molecules. Meeting regulatory standards necessitates large quantities of biotherapeutic agents of high purity. While conventional chromatography offers versatility and efficiency, it suffers from drawbacks like low flow rates and binding capacity, as well as high mass transfer resistance. Recent advancements in continuous beds, including monoliths and cryogel-based systems, have emerged as promising solutions to overcome these limitations. This review explores and evaluates the latest progress in chromatography utilizing monolithic and cryogenic supports for nucleic acid purification.

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Section: Cryogelsmentioning

confidence: 99%

Section: Dna Purificationmentioning

confidence: 99%

Cryogels and Monoliths: Promising Tools for Chromatographic Purification of Nucleic Acids

Ribeiro,

Luís,

Rodrigues

et al. 2024

Gels

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“…According to the mathematical Langmuir model, the column has shown monolayer adsorption kinetics. Moreover, the column was found intact without deformation in shape post 30 times adsorption and desorption cycles (Önal & Odabas ¸ı, 2021). Despite the good performance and reusability, in IMAC other biomolecules from the crude lysate can form the complexes with the metal that reduces the specificity of the column.…”

Section: Immobilized Metal Affinity Chromatographymentioning

confidence: 99%

Functionalized cryogel monoliths for fast and selective separation of nucleic acids directly from crude lysate

Shakya

2022

Biomedical Chromatography

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The fast and selective separation of nucleic acids has been attractive recently because of their wide number of applications in the biomedical field such as the development of vaccines for infectious diseases, gene therapy, and diagnosis. Traditional approaches of nucleic acids separation are costlier, lengthy, and associated with possible denaturation because of the use of organic solvents in the elution step.

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“…Cryogels have 3D elastic interconnected macropores, spongy morphologies and good mechanical robustness [3]. They are extremely versatile tools when converted to desired purposes [4] such as purification/adsorption of biomolecules [5][6][7][8], and removal of some pollutants [9][10][11]. Increasing the existing properties while designing cryogels will further increase their potential use in different fields [4].…”

Section: Introductionmentioning

confidence: 99%

Immobilization of xylanase enzyme on poly-(HEMA-co-GMA) cryogel

Noma

2022

MANAS Journal of Engineering

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In this study, a polyethyleneimine (PEI) coated poly-(HEMA-co-GMA), hybrid cryogel column (HCC), was designed. HCC was synthesized via polymerization of gel-former factors at minus temperatures. The characterization experiments of the HCC were conducted through SEM, and FTIR experiments. At the end of the experimental periods, there was no significant decrease in the performance of the HCC. Then the HCC was used as a novel support for xylanase immobilization for the first time. the successful immobilization of xylanase was confirmed by FT-IR, while the biochemical properties and stability of the PHG/PI-Xyl were evaluated in terms of optimum pH, optimum temperature, thermostability, storage stability, reusability, and kinetic parameters, The optimum activities for both free and immobilized enzyme were recorded at pH 6.0, while optimum temperature for free was 55 °C and for PHG/PI-Xyl 60 °C. The PHG/PI-Xyl displayed remarkable thermal stability for 180 min at 60 °C, with 53.55%, and for free Xyl 32.05%. Meanwhile, it retained up to 49% and 69 % for free and immobilized xylanase of original activities after 4 weeks of storage at room temperature. The PHG/PI-Xyl retained about 58% of its original activity after 10 consecutive reuses, while Km for the free Xyl and PHG/PI-Xyl were calculated at 4.05 mg/mL and 2.62 mg/mL, whereas Vmax133.33 U/mL and 188.68 U/mL, respectively. As envisioned, this study suggests a promising way to solve the problems of high price and poor operational stability of the enzyme during biocatalytic.

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Design and application of a newly generated bio/synthetic cryogel column for DNA capturing

Cited by 8 publications

References 61 publications

Cryogels and Monoliths: Promising Tools for Chromatographic Purification of Nucleic Acids

Cryogels and Monoliths: Promising Tools for Chromatographic Purification of Nucleic Acids

Functionalized cryogel monoliths for fast and selective separation of nucleic acids directly from crude lysate

Immobilization of xylanase enzyme on poly-(HEMA-co-GMA) cryogel

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