Adsorption of deamidated antibody variants on macroporous and dextran-grafted cation exchangers: I. Adsorption equilibrium

Tao, Yinying; Carta, Giorgio; Ferreira, Gisela; Robbins, David J.

doi:10.1016/j.chroma.2011.01.049

Cited by 59 publications

(32 citation statements)

References 25 publications

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“…In this status, the ligands immobilized on dextran-grafted MMC adsorbents were stuck together via the hydrophobic interaction, consequently resulting in the collapse of dextran layer in MMC adsorbent. Tao et al also found the partial collapse of dextran polymer at high ion strength as a result of shielding of repulsive intra-and interchain electrostatic interactions [41]. Therefore, at pH 8.0, the pore architecture in dextran-grafted MMC adsorbent did not change obviously with the grafting of dextran and surface area was still the limiting factor for protein adsorption.…”

Section: The Effect Of Dextran Contentmentioning

confidence: 95%

“…In contrast to MMC adsorbents, ionic group bound to the grafted dextran in the pores of IEC adsorbents repelled each other, hence leading to the formation of the inflated polymer layer throughout the pH range used herein [42]. At higher ion strength, the inflated polymer layer could partially collapse [41]. On the other hand, e p for g-globulin in Table 2 showed that MMC adsorbents had the same value of e p as IEC adsorbent as the dextran content is constant.…”

Section: The Effect Of Dextran Contentmentioning

confidence: 95%

“…It showed an obvious increase in adsorption capacities of g-globulin on dextran-grafted SP-T40-X-S6B and SP Sepharose XL compared with those on non-grafted IEC adsorbents. 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]).…”

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: The Effect Of Dextran Contentmentioning

confidence: 95%

Section: The Effect Of Dextran Contentmentioning

confidence: 95%

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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“…Nevertheless, the complete size exclusion of the 40 kDa dextran suggests a pore radius of about 5 nm, Curve labels give dextran molecular mass and hydrodynamic radius estimated as the average values given by Squire 25 and Hagel et al 26 similar to the hydrodynamic radius of IgG ($5 nm), but larger than the hydrodynamic radius of lysozyme ($2 nm). By comparison, UNOsphere S has a mean pore radius around 68 nm, 28 much larger than the hydrodynamic radius of either test protein used in this work. Figure 5 shows the adsorption isotherms for lysozyme and IgG in phosphate buffer at pH 6.5 and in acetate buffer at pH 5.0, respectively.…”

mentioning

confidence: 96%

Protein adsorption and transport in cation exchangers with a rigid backbone matrix with and without polymeric surface extenders

Almodovar

Tao

Carta

2011

Biotechnology Progress

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We compare the properties and protein adsorption characteristics of two polymeric cation exchangers: UNOsphere S, which has an open macroporous architecture, and Nuvia S, which is based on a very similar backbone matrix but contains sulfonated polymeric surface extenders. A monoclonal IgG and lysozyme were used as model adsorbates. The characteristic pore sizes, determined by inverse size exclusion chromatography, were about 140 nm for UNOsphere S, and only about 10 nm for Nuvia S, indicating that the polymeric extenders occupy a substantial portion of the base matrix pores. Greater exclusion limits were found for Nuvia S in 1 M NaCl and for a similar matrix containing uncharged surface extenders, suggesting that the polymeric extenders collapse partially at high ionic strength or when they are uncharged. Large equilibrium binding capacities were obtained for Nuvia S, approaching 320 ± 10 mg/mL of particle volume for both proteins in comparison with the UNOsphere S values of 170 ± 10 and 120 ± 10 mg/mL for lysozyme and IgG, respectively. Much higher adsorption rates were also found for Nuvia S, and the rate was nearly independent of protein concentration in solution. Confocal laser scanning microscopy showed very sharp intraparticle protein concentration profiles for UNOsphere S, consistent with a pore diffusion mechanism but diffuse concentration profiles for Nuvia S, consistent with a solid diffusion mechanism. The improved capacity and transport afforded by the polymeric extenders provide substantial potential benefits for bioprocess applications without sacrificing the desirable flow properties of the backbone matrix.

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“…Polymer-grafted adsorbent has been researched extensively in ion exchange chromatography (IEC) in recent years [1][2][3][4]. For example, as the representative polymer-grafted ion-exchangers, dextran-grafted IEC resins [2,5], such as the commercial SP Sepharose XL, Streamline Q XL [2,[6][7][8], and the customized T40-SP-X-S6B [9] were widely used for antibody purification.…”

Section: Introductionmentioning

confidence: 99%

Implications from protein uptake kinetics onto dextran-grafted Sepharose FF coupled with ion exchange and affinity ligands

Xue

Sun

2017

Chinese Journal of Chemical Engineering

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Adsorption of deamidated antibody variants on macroporous and dextran-grafted cation exchangers: I. Adsorption equilibrium

Cited by 59 publications

References 25 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

Protein adsorption and transport in cation exchangers with a rigid backbone matrix with and without polymeric surface extenders

Implications from protein uptake kinetics onto dextran-grafted Sepharose FF coupled with ion exchange and affinity ligands

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