Preparation and characterization of grafted cellulosic fibers and their applications in protein purification

Singh, Naveen Kumar; Dsouza, Roy N.; Sánchez, María Amparo; Verma, Sujit Kumar; Achilli, Estefanía Edith; Vennapusa, Rami Reddy; Grasselli, Mariano; Fernández‐Lahore, Marcelo

doi:10.1016/j.seppur.2015.01.042

Cited by 9 publications

(11 citation statements)

References 50 publications

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“…Grafting highly porous materials can improve protein binding capacity by generating either an active “gel” phase or by creating a polymer “brush”. This possibility was proven in previous work performed by us .…”

Section: Resultssupporting

confidence: 70%

A megaporous material harbouring a peptide ligand for affinity IgG purification

et al. 2017

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Common limitations of Protein A affinity chromatography include high adsorbent costs, ligand instability and possible ligand leakage. In this study, a short peptide with affinity for IgG was synthesized chemically and subsequently immobilized on a megaporous support. The support was prepared utilising the cryogel technique while the peptide-ligand was covalently immobilised via thiol-epoxy click chemistry. The cryogel support was chemically grafted to increase the number of reaction sites. This adsorbent was designated as "MP-Pep". Adsorption isotherms were employed to evaluate protein binding capacity. A maximum static binding capacity within the range of 30-60 mg/mL was observed for hIgG. This parameter compares well with other commercial and non-commercial adsorbents, as reported in the literature. As a control material, a Protein A grafted megaporous cryogel was synthesized. Dynamic binding capacity values were obtained by breakthrough analysis. The peptide cryogel showed a dynamic capacity value 9.0 mg/mL in comparison to 9.7 mg/mL in the case of the Protein A based adsorbent. The ratio of dynamic binding capacity to static binding capacity was 20%, indicating suboptimal product capture. However, the advantage of MP-Pep lies in its cost-effective preparations while maintaining a reasonable binding capacity for the targeted product. The presence of cooperative effects during protein binding could also represent an advantage during the processing of a feedstock containing a product in high concentration.

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Section: Resultssupporting

confidence: 70%

A megaporous material harbouring a peptide ligand for affinity IgG purification

et al. 2017

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“…Later on, using a similar approach, chemical grafting was also performed on the alkaline-treated cotton fibres for preparing the chromatographic materials. 15 Composite fibres containing different ionic ligands, such as ionexchange fibres, were successfully prepared. They showed very high protein adsorption capacity when they were loaded in standard chromatographic columns.…”

Section: Resultsmentioning

confidence: 99%

“…Fibres were pretreated with an alkaline solution, as described in Singh. 15 After exhaustive washing, they were dried and used for further functionalization. Composite fibres were prepared by radiation-induced grafting polymerisation.…”

Section: Composite Preparationmentioning

confidence: 99%

Composite Cellulose Fibre for Affinity Chromatography Application

Carbajal¹,

Kikot²,

Torchio³

et al. 2020

Cellulose Chem. Technol.

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Affinity chromatographic supports are nowadays one of the most frequently used and expensive consumable materials for protein purification at the laboratory and industrial scales. The introduction of cost-effective materials is an important issue to address in order to spread the usefulness of this technology. Cotton fibres are a highly available natural material with excellent mechanical and structural properties, which can be used for this purpose. Nevertheless, fibre insolubility and low chemical reactivity are the major drawbacks preventing the use of this material for protein chromatography. In this work, a composite prepared from polymethacrylate/cellulose fibres was used for the preparation of chromatographic materials containing immobilised proteins as adsorptive ligands. A green fluorescent protein and a protein A-similar ligand, containing cysteine in the terminal tags, were immobilised onto the fibres by epoxy-thiol chemistry. Buffer salt, pH, reaction time and pre-swelling procedure were optimised. The materials were characterised by fluorescence and electron microscopy, in addition to the binding capacity of the adsorbent materials.

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“…This was performed by grafting with a single vinyl‐containing monomer such as GMA and two modes of initiators, physical and chemical. Finally, pendant epoxy groups were modified to provide weak anion‐exchange functionality . The (dynamic) binding capacity of the composite anion exchange, high voidage, adsorptive systems was investigated for pDNA purification.…”

Section: Resultsmentioning

confidence: 99%

“…The CG‐DEAE‐fibers and GIR‐DEAE cryogels were sythesized as previously reported . They were packed in 5.5 cm × 5 mm and 3.4 cm × 10 mm columns, respectively (GE Amersham Biosciences, Uppsala, Sweden) and connected to a Äkta Explorer 100 having UNICORN™ 4.10 software (GE Amersham Biosciences).…”

Section: Methodsmentioning

confidence: 99%

pDNA capture using grafted adsorbents

Singh

Dsouza

Yelemane

et al. 2018

J of Chemical Tech & Biotech

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BACKGROUND: 'Expanded' composite materials are of interest as an alternative, or as a supplement, to packed-bed chromatography during bioproduct recovery and purification. Functionalized non-woven fabrics and mega-porous bodies are examples of systems that showed promise. However, there is scarce information on their suitability to capture and release plasmid DNA (pDNA), an important type of product employed in gene therapy. RESULTS: Composite adsorbents were prepared using either chemical (CG-DEAE-NW) or gamma-irradiated graft-polymerization (GIR-DEAE-MP), and subsequently modified to have diethylamino ethanol (DEAE) functionality. Capture experiments showed that pDNA can actually reversibly bind to the two mentioned adsorbents, with capacity values of 2.4 and 1.3 mg per mL, respectively. These values are in the range of what can be expected from commercial beaded adsorbents but lower that the values expected from monoliths. CONCLUSIONS: Expanded materials, due to their high voidage, may present limited capacity for pDNA. However, such materials are able to bind proteins and other contaminants from bacterial lysate, opening the way for their utilization in the 'negative' mode.

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Preparation and characterization of grafted cellulosic fibers and their applications in protein purification

Cited by 9 publications

References 50 publications

A megaporous material harbouring a peptide ligand for affinity IgG purification

A megaporous material harbouring a peptide ligand for affinity IgG purification

Composite Cellulose Fibre for Affinity Chromatography Application

pDNA capture using grafted adsorbents

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