Comparative study of strong anion exchangers: Structure-related chromatographic performances

Pezzini, J.; Cabanne, Charlotte; Santarelli, Xavier

doi:10.1016/j.jchromb.2009.06.044

Cited by 10 publications

(1 citation statement)

References 20 publications

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“…Using three common AEX resins with widely different structures, the specific goal of this work is to investigate the partitioning and mass transfer behavior of mAb monomeric species along with that of (a) mAb dimers and mAb multimeric species, and (b) BSA as a model of typical acidic impurities. 17 The three resins considered are the tentacle-type, polymethacrylate-based Fractogel V R EMD TMAE HiCap (M) from EMD Millipore (Billerica, MA), hereinafter referred to as TMAE, the relatively small-pore, agarose-based Q Sepharose FF from GE Healthcare (Piscataway, NJ), hereinafter referred to as QFF, and the large-pore, polystyrene-based POROS 50 HQ from Thermo Fisher Scientific (Waltham, MA), hereinafter referred to as POROS. The structure of the three resins is determined by transmission electron microscopy, their pore accessibility by inverse size exclusion chromatography, and their charge density by potentiometric titrations.…”

Section: Introductionmentioning

confidence: 99%

Structural and performance characteristics of representative anion exchange resins used for weak partitioning chromatography

Zhang

Iskra

Daniels

et al. 2017

Biotechnology Progress

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Weak partitioning chromatography (WPC) has been proposed for the purification of monoclonal antibodies using an anion exchange (AEX) resin to simultaneously remove both acidic and basic protein impurities. Despite potential advantages, the relationship between resin structure and WPC performance has not been evaluated systematically. In this work, we determine the structure of representative AEX resins (Fractogel® EMD TMAE HiCap, Q Sepharose FF, and POROS 50 HQ) using transmission electron microscopy and inverse size exclusion chromatography and characterize protein interactions while operating these resins under WPC conditions using two mAb monomers, a mAb dimer, mAb multimers, and BSA as model products and impurities. We determine the isocratic elution behavior of the weakly bound monomer and dimer species and the adsorptive and mass transfer properties of the strongly bound multimers and BSA by confocal laser scanning microscopy. The results show that for each resin, using the product K value as guidance, salt, and pH conditions can be found where mAb multimers and BSA are simultaneously removed. Isocratic elution and adsorption mechanisms are, however, different for each resin and for the different components. Under WPC conditions, the Fractogel resin exhibited very slow diffusion of both mAb monomer and dimer species but fast adsorption for both mAb multimers and BSA with high capacity for BSA, while the Sepharose resin, because of its small pore size, was unable to effectively remove mAb multimers. The POROS resin was instead able to bind both multimers and BSA effectively, while exhibiting a greater resolution of mAb monomer and dimer species. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:425-434, 2017.

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Section: Introductionmentioning

confidence: 99%

Structural and performance characteristics of representative anion exchange resins used for weak partitioning chromatography

Zhang

Iskra

Daniels

et al. 2017

Biotechnology Progress

View full text Add to dashboard Cite

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Exploring the use of heparin as a first capture step in the purification of monoclonal antibodies from cell culture supernatants

Pinto

Rosa

Aires-Barros

et al. 2015

Biochemical Engineering Journal

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Protein adsorption and transport in polymer-functionalized ion-exchangers

Lenhoff

2011

Journal of Chromatography A

115

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A wide variety of stationary phases is available for use in preparative chromatography of proteins, covering different base matrices, pore structures and modes of chromatography. There has recently been significant growth in the number of such materials in which the base matrix is derivatized to add a covalently attached or grafted polymer layer or, in some cases, a hydrogel that fills the pore space. This review summarizes the main structural and functional features of ion exchangers of this kind, which represent the largest class of such materials. Although the adsorption and transport properties may generally be used operationally and modeled phenomenologically using the same methods as are used for proteins in conventional media, there are noteworthy mechanistic differences in protein behavior in these adsorbents. A fundamental difference in protein retention is that it may be portrayed as partitioning into a three-dimensional polymer phase rather than adsorption at an extended two-dimensional surface, as applies in more conventional media. Beyond this partitioning behavior, however, the polymer-functionalized media often display rapid intraparticle transport that, while qualitatively comparable to that in conventional media, is sufficiently rapid quantitatively under certain conditions that it can lead to clear benefits in key measures of performance such as the dynamic binding capacity. Although possible mechanistic bases for the retention and transport properties are discussed, appreciable areas of uncertainty make detailed mechanistic modeling very challenging, and more detailed experimental characterization is likely to be more productive.

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Comparative study of strong anion exchangers: Structure-related chromatographic performances

Cited by 10 publications

References 20 publications

Structural and performance characteristics of representative anion exchange resins used for weak partitioning chromatography

Structural and performance characteristics of representative anion exchange resins used for weak partitioning chromatography

Exploring the use of heparin as a first capture step in the purification of monoclonal antibodies from cell culture supernatants

Protein adsorption and transport in polymer-functionalized ion-exchangers

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