Properties and Characterization of High Capacity Resins for Biochromatography

Müller, Egbert

doi:10.1002/ceat.200500161

Cited by 84 publications

(59 citation statements)

References 50 publications

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“…Therefore, it was speculated that the grafting of dextran led to peculiar pore architecture in dextran-grafted IEC adsorbent and provided greater accessibility for the protein [8,23,32,41]. Compared with ligand immobilized on pore surface in SP-SEP, IEC ligand uniformly distributed in grafted-dextran layer above pore surface in dextran-grafted IEC adsorbents (SP-D1-SEP, SP-D2-SEP and SP-D3-SEP), exhibiting a three-dimensional orientation of ligand molecules (a pictorial presentation in [42]). Therefore, protein molecules could adsorb along the functionalized dextran.…”

Section: Adsorption Isothermsmentioning

confidence: 98%

“…As one of the most important determinants for the performance of protein chromatography, adsorption capacity for proteins depended on the property of adsorbents, particularly on the surface area, pore volume or available charge of the adsorbent [15,42,45]. For a typical IEC adsorbent, the charged ligands were tightly stacked to provide excessive sites for adsorption, and therefore, surface area or pore volume was considered as the limiting factor for protein adsorption [15].…”

Section: Adsorption Isothermsmentioning

confidence: 99%

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Effect of dextran layer on protein uptake to dextran‐grafted adsorbents for ion‐exchange and mixed‐mode chromatography

Shi

Sun

2011

J of Separation Science

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In the current research, a series of dextran-grafted adsorbents were prepared using sulfopropyl and 4-(1H-imidazol-1-yl) aniline as chromatographic ligands for ion-exchange (IEC) and mixed-mode chromatography (MMC) to respectively investigate the influence of dextran layer on adsorption of γ-globulin. Experimental evidences of static adsorption on dextran-grafted IEC adsorbents showed that adsorption capacity of γ-globulin increased with dextran content. It could be attributed to the multilayer adsorption of charged protein in dextran layer and thus further induced a significant electrical potential gradient at the boundary of adsorbed area and its proximity, improving mass transfer in combination with concentration gradient. In contrast to IEC adsorbents, adsorption capacity and effective diffusivity of dextran-grafted MMC adsorbents did not change obviously with dextran grafting. It was considered that hydrophobic ligands immobilized onto dextran-grafted MMC adsorbents were stuck together at pH 8.0, resulting in the collapse of dextran layer. In concert with measured effective porosity for γ-globulin at pH 4.0, it was confirmed that dextran layer in MMC adsorbent was more complicated and influenced significantly by buffer pH. It was also manifested by protein adsorption at different pHs. Thus, it revealed the complexity in intraparticle mass transfer of the protein in dextran-grafted MMC adsorbent.

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Section: Adsorption Isothermsmentioning

confidence: 98%

Section: Adsorption Isothermsmentioning

confidence: 99%

Effect of dextran layer on protein uptake to dextran‐grafted adsorbents for ion‐exchange and mixed‐mode chromatography

Shi

Sun

2011

J of Separation Science

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“…Indeed, a 70% higher value of DBC over EBC (2.18 vs. determined under practically relevant conditions can be 5-10 times less than the EBCs for a given phase. 63,64 To be clear, under flow conditions, there would be lesser utilization of any porosity as those diffusional processes would be kinetically limited. In fact, a value of unity for DBC/EBC could only be achieved under the situation of infinitely high chromatographic efficiencies and rapid mass transfer kinetics.…”

Section: Role Of Fiber Packing Density At Fixed Volume Flowmentioning

confidence: 99%

Roles of interstitial fraction and load conditions on the dynamic binding capacity of proteins on capillary‐channeled polymer fiber columns

Wang

Marcus

2014

Biotechnology Progress

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Capillary-channeled polymer (C-CP) fibers are used as a stationary phase for ion-exchange chromatography of proteins. Collinear packing of the fibers permits operation at high linear velocities (Uo > 100 mm s(-1)) and low backpressure (<2,000 psi) on analytical-scale columns. Rapid solvent transport is matched with very efficient solute mass transfer as fibers are virtually non-porous with respect to the size of the target protein molecules. Lack of porosity of course limits the equilibrium binding capacity of stationary phases. Breakthrough curves and frontal analysis are used to better understand trade-offs between the kinetic and thermodynamic properties as C-CP fibers are applied in preparative situations. Fiber columns packed to different interstitial fraction values affect both the total fiber surface area (e.g., equilibrium binding capacity [EBC]) and the permittivity to flow and mass transport characteristics (e.g., dynamic binding capacity [DBC]). The EBC of the nylon 6 C-CP fibers was found to be 1.30 mg g(-1), with isotherms that were best matched by a Moreau model, showing linearity up to solute concentrations of ∼0.4 mg mL(-1). Isotherms generated under flow conditions were equally well approximated using Langmuir, Freundlich, and Moreau isotherm models. Fairly linear responses were seen up to the maximum load concentration of 1.2 mg mL(-1). Counterintuitively, dynamic studies revealed that conditions of high column porosity yielded a DBC that is ∼70% higher than the EBC. These findings point to potential advantages in terms downstream processing applications, where protein throughput and yield are critical metrics.

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“…Commercial butylated chromato- graphic adsorbents offer adsorption capacities varying from 27 to 39 mg BSA/mL medium, as obtained from GE Healthcare data sheet products, under dynamic conditions. Normally, lower residence times decrease the DBC [21], while longer residence times lead to higher adsorption capacities as the protein molecules have more time to travel to interior of the pores of the resin beads and interact to a great extent with ligands [22]. Interestingly, in this work the DBC was higher as the flow rate increased.…”

Section: Static and Dynamic Adsorption Capacitiesmentioning

confidence: 66%

Synthesis of adsorbents with dendronic structures for protein hydrophobic interaction chromatography

Mata-Gómez

Yaman

Valencia-Gallegos

et al. 2016

Journal of Chromatography A

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a b s t r a c tHere, we introduced a new technology based on the incorporation of dendrons-branched chemical structures-onto supports for synthesis of HIC adsorbents. In doing so we studied the synthesis and performance of these novel HIC dendron-based adsorbents. The adsorbents were synthesized in a facile two-step reaction. First, Sepharose 4FF (R) was chemically modified with polyester dendrons of different branching degrees i.e. third (G3) or fifth (G5) generations. Then, butyl-end valeric acid ligands were coupled to dendrons via ester bond formation. UV-vis spectrophotometry and FTIR analyses of the modified resins confirmed the presence of the dendrons and their ligands on them. Inclusion of dendrons allowed the increment of ligand density, 82.5 ± 11 and 175.6 ± 5.7 mol ligand/mL resin for RG3 and RG5, respectively. Static adsorption capacity of modified resins was found to be ∼60 mg BSA/mL resin. Interestingly, dynamic binding capacity was higher at high flow rates, 62.5 ± 0.8 and 58.0 ± 0.5 mg/mL for RG3 and RG5, respectively. RG3 was able to separate lipase, ␤-lactoglobulin and ␣-chymotrypsin selectively as well as fractionating of a whole proteome from yeast. This innovative technology will improve the existing HIC resin synthesis methods. It will also allow the reduction of the amount of adsorbent used in a chromatographic procedure and thus permit the use of smaller columns resulting in faster processes. Furthermore, this method could potentially be considered as a green technology since both, dendrons and ligands, are formed by ester bonds that are more biodegradable allowing the disposal of used resin waste in a more ecofriendly manner when compared to other exiting resins.

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Properties and Characterization of High Capacity Resins for Biochromatography

Cited by 84 publications

References 50 publications

Effect of dextran layer on protein uptake to dextran‐grafted adsorbents for ion‐exchange and mixed‐mode chromatography

Effect of dextran layer on protein uptake to dextran‐grafted adsorbents for ion‐exchange and mixed‐mode chromatography

Roles of interstitial fraction and load conditions on the dynamic binding capacity of proteins on capillary‐channeled polymer fiber columns

Synthesis of adsorbents with dendronic structures for protein hydrophobic interaction chromatography

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