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

Shakya, Akhilesh Kumar

doi:10.1002/bmc.5333

Cited by 6 publications

(2 citation statements)

References 37 publications

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“…Cryogels ensure unrestrictive convective transport, whereas the latter can only perform mass transport through diffusion [ 92 ]. As a result, cryogels facilitate the efficient separation of nanoparticles, cellular organelles, and even whole cells via chromatographic methods [ 93 ]. The convective flow, as depicted in Figure 2 , is exclusively facilitated by the macropores inherent in the cryogels, ranging in size from 10 to 100 µm [ 94 ].…”

Section: Cryogelsmentioning

confidence: 99%

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%

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

Ribeiro,

Luís,

Rodrigues

et al. 2024

Gels

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“…Apart from strengthening the boronate affinity, overcoming the hindrance of interfering matrix in bioseparation is another key point for bacterial analysis . 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.…”

Section: Introductionmentioning

confidence: 99%

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

Yan,

Wei,

Gou

et al. 2024

J. Agric. Food Chem.

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The lack of practical platforms for bacterial separation remains a hindrance to the detection of bacteria in complex samples. Herein, a composite cryogel was synthesized by using clickable building blocks and boronic acid for bacterial separation. Macroporous cryogels were synthesized by cryo-gelation polymerization using 2-hydroxyethyl methacrylate and allyl glycidyl ether. The interconnected macroporous architecture enabled high interfering substance tolerance. Nanohybrid nanoparticles were prepared via surface-initiated atom transfer radical polymerization and immobilized onto cryogel by click reaction. Alkyne-tagged boronic acid was conjugated to the composite for specific bacteria binding. The physical and chemical characteristics of the composite cryogel were analyzed systematically. Benefitting from the synergistic, multiple binding sites provided by the silica-assisted polymer, the composite cryogel exhibited excellent affinity toward S. aureus and Salmonella spp. with capacities of 91.6 × 10 7 CFU/g and 241.3 × 10 7 CFU/g in 0.01 M PBS (pH 8.0), respectively. Bacterial binding can be tuned by variations in pH and temperature and the addition of monosaccharides. The composite was employed to separate S. aureus and Salmonella spp. from spiked tap water, 40% cow milk, and sea cucumber enzymatic hydrolysate, which resulted in high bacteria separation and demonstrated remarkable potential in bacteria separation from food samples.

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Specific DNA aptamer–immobilized cryogel membranes as novel bioaffinity supports and their potential for the purification of activated protein C

Aliyeva,

Akgönüllü,

Erdem

et al. 2024

Biomedical Chromatography

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Activated protein C (APC), a serine protease produced from zymogen protein C (PC), is the key enzyme of the protein C pathway. APC has anticoagulant, anti‐inflammatory, and cytoprotective features. APC has recently been shown to significantly reduce coagulation as well as mortality in patients with severe sepsis. Herein, we aimed to develop an affinity support material that allows the purification of plasma APC for the first time. In this research, a novel APC‐specific DNA aptamer–based poly(2‐hydroxyethyl methacrylate‐glycidyl methacrylate) (poly(HEMA‐GMA/DNA‐Apt)) macroporous cryogel membrane at different molar ratios was prepared using affinity binding method and their potential for purification and identification of APC was investigated. The DNA aptamer–immobilized cryogels were characterized to examine their structural and morphological properties. The effect of pH, initial concentration, temperature, ionic strength difference, and flow rate changes was examined. Selectivity studies were performed in the presence of APC and competitive proteins, and cryogel support materials were shown to have a very high affinity for APC. Adsorption capacity was found to be 89.02 mg/g. Finally, NaCl revealed efficiency for APC desorption and the reuse of cryogels was successfully tested for ten cycles.

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

Cited by 6 publications

References 37 publications

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

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

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

Specific DNA aptamer–immobilized cryogel membranes as novel bioaffinity supports and their potential for the purification of activated protein C

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