Protein adsorption and transport in polymer-functionalized ion-exchangers.

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.

[1]  R. Carbonell,et al.  The adsorption of proteins to gas-liquid interfaces , 1986 .

[2]  E. Müller Properties and Characterization of High Capacity Resins for Biochromatography , 2005 .

[3]  G. Carta,et al.  Protein adsorption on novel acrylamido-based polymeric ion-exchangers. III. Salt concentration effects and elution behavior. , 2001, Journal of chromatography. A.

[4]  Duncan Low,et al.  Future of antibody purification. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[5]  C. Cabanne,et al.  Comparative study of strong anion exchangers: structure-related chromatographic performances. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[6]  A. Lenhoff,et al.  A predictive approach to correlating protein adsorption isotherms on ion-exchange media. , 2008, The journal of physical chemistry. B.

[7]  P. Dephillips,et al.  Pore size distributions of cation-exchange adsorbents determined by inverse size-exclusion chromatography. , 2000, Journal of chromatography. A.

[8]  Uwe Gottschalk,et al.  Bioseparation in Antibody Manufacturing: The Good, The Bad and The Ugly , 2008, Biotechnology progress.

[9]  H. Bysell,et al.  Biomacromolecules in microgels — Opportunities and challenges for drug delivery , 2010 .

[10]  T. Przybycien,et al.  Alternative bioseparation operations: life beyond packed-bed chromatography. , 2004, Current opinion in biotechnology.

[11]  G. Guiochon,et al.  Study of the physico-chemical properties of some packing materials. III. Pore size and surface area distribution , 1997 .

[12]  Abraham M Lenhoff,et al.  Protein adsorption and transport in dextran-modified ion-exchange media. II. Intraparticle uptake and column breakthrough. , 2011, Journal of chromatography. A.

[13]  A. Lenhoff,et al.  Comparison of protein adsorption isotherms and uptake rates in preparative cation-exchange materials. , 1998, Journal of chromatography. A.

[14]  Arne Staby,et al.  Comparison of chromatographic ion-exchange resins V. Strong and weak cation-exchange resins. , 2006, Journal of chromatography. A.

[15]  A. Dobrynin,et al.  Theory of polyelectrolytes in solutions and at surfaces , 2005 .

[16]  John E. Sader,et al.  Electrostatic Contribution to the Energy and Entropy of Protein Adsorption , 1998 .

[17]  Brian Kelley,et al.  Very Large Scale Monoclonal Antibody Purification: The Case for Conventional Unit Operations , 2007, Biotechnology progress.

[18]  Motonobu Yoshikawa,et al.  Parallel transport of BSA by surface and pore diffusion in strongly basic chitosan , 1994 .

[19]  G. Carta,et al.  Protein partitioning and transport in supported cationic acrylamide-based hydrogels , 2003 .

[20]  G. Carta,et al.  Protein adsorption on novel acrylamido-based polymeric ion-exchangers. I. Morphology and equilibrium adsorption. , 2000, Journal of chromatography. A.

[21]  F. Regnier,et al.  Retention model for high-performance ion-exchange chromatography☆ , 1983 .

[22]  D. Ruthven,et al.  Adsorption of water from aqueous ethanol using 3-.ANG. molecular sieves , 1986 .

[23]  G. Carta,et al.  Protein Transport in Constrained Anionic Hydrogels: Diffusion and Boundary-Layer Mass Transfer , 2001 .

[24]  G. Carta,et al.  Rapid monoclonal antibody adsorption on dextran-grafted agarose media for ion-exchange chromatography. , 2008, Journal of chromatography. A.

[25]  C. Roth,et al.  Electrostatic and van der Waals Contributions to Protein Adsorption: Comparison of Theory and Experiment , 1995 .

[26]  M. Tirrell,et al.  Functional polymer brushes in aqueous media from self-assembled and surface-initiated polymers. , 2008, Annual review of physical chemistry.

[27]  P. Levison,et al.  Performance comparison of low-pressure ion-exchange chromatography media for protein separation , 1997 .

[28]  J. A. Wesselingh,et al.  Protein ion-exchange adsorption kinetics , 2001 .

[29]  Giorgio Carta,et al.  Characterization of protein adsorption by composite silica-polyacrylamide gel anion exchangers II. Mass transfer in packed columns and predictability of breakthrough behavior , 1996 .

[30]  P. Hansson,et al.  Distribution of cytochrome c in polyacrylate microgels , 2010 .

[31]  J. V. Van Alstine,et al.  Ion exchange chromatography of antibody fragments , 2007, Biotechnology and bioengineering.

[32]  Giorgio Carta,et al.  Protein Mass Transfer Kinetics in Ion Exchange Media: Measurements and Interpretations , 2005 .

[33]  G. Carta,et al.  Characterization of protein adsorption by composite silica-polyacrylamide gel anion exchangers I. Equilibrium and mass transfer in agitated contactors , 1996 .

[34]  J. V. Van Alstine,et al.  Modeling of protein interactions with surface-grafted charged polymers. Correlations between statistical molecular modeling and a mean field approach. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[35]  P. Krajnc,et al.  Characterisation of grafted weak anion-exchange methacrylate monoliths. , 2008, Journal of chromatography. A.

[36]  Massimo Morbidelli,et al.  Electrostatic model for protein adsorption in ion-exchange chromatography and application to monoclonal antibodies, lysozyme and chymotrypsinogen A. , 2010, Journal of chromatography. A.

[37]  Frank G. Smith,et al.  Electrostatic effects on the partitioning of spherical colloids between dilute bulk solution and cylindrical pores , 1983 .

[38]  J. V. Van Alstine,et al.  An exclusion mechanism in ion exchange chromatography. , 2006, Biotechnology and bioengineering.

[39]  W. Deen Hindered transport of large molecules in liquid‐filled pores , 1987 .

[40]  Bernard Sebille,et al.  Modeling of Protein Adsorption on Polymer Surfaces. Computation of Adsorption Potential , 1995 .

[41]  G. Malmquist,et al.  Surface extenders and an optimal pore size promote high dynamic binding capacities of antibodies on cation exchange resins. , 2009, Journal of chromatography. A.

[42]  P. Dephillips,et al.  Determinants of protein retention characteristics on cation-exchange adsorbents. , 2001, Journal of chromatography. A.

[43]  Ryuichi Matsuno,et al.  Ion-Exchange Chromatography of Proteins , 1988 .

[44]  A. Lenhoff,et al.  Nondiffusive mechanisms enhance protein uptake rates in ion exchange particles , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[45]  Abraham M Lenhoff,et al.  Electrostatic contributions to protein retention in ion-exchange chromatography. 1. Cytochrome C variants. , 2004, Analytical chemistry.

[46]  J. Israelachvili Intermolecular and surface forces , 1985 .

[47]  F. Švec,et al.  Monolithic materials : preparation, properties and applications , 2003 .

[48]  A. Staby,et al.  Comparison of chromatographic ion-exchange resins , 2000 .

[49]  Abraham M. Lenhoff,et al.  Influence of Structural Details in Modeling Electrostatically Driven Protein Adsorption , 1997 .

[50]  K. Sing,et al.  The use of nitrogen adsorption for the characterisation of porous materials , 2001 .

[51]  A. Zydney,et al.  Modeling electrostatic exclusion effects during ion exchange chromatography of monoclonal antibodies , 2009, Biotechnology and bioengineering.

[52]  M. Kula,et al.  Dynamics of protein uptake within the adsorbent particle during packed bed chromatography. , 2002, Biotechnology and bioengineering.

[53]  C. L. Y. Leon,et al.  New perspectives in mercury porosimetry , 1998 .

[54]  Giorgio Carta,et al.  Protein adsorption and transport in agarose and dextran-grafted agarose media for ion exchange chromatography: Effect of ionic strength and protein characteristics. , 2009, Journal of chromatography. A.

[55]  A. Staby,et al.  Comparison of chromatographic ion-exchange resins IV. Strong and weak cation-exchange resins and heparin resins. , 2005, Journal of chromatography. A.

[56]  G. Carta,et al.  Protein Adsorption on Cation Exchangers: Comparison of Macroporous and Gel‐Composite Media , 1996 .

[57]  J. Janson Protein purification: Principles, high resolution methods, and applications , 1989 .

[58]  A. Lenhoff,et al.  Chromatography of proteins on charge-variant ion exchangers and implications for optimizing protein uptake rates. , 2007, Journal of chromatography. A.

[59]  A. Khademhosseini,et al.  Hydrogels in Regenerative Medicine , 2009, Advanced materials.

[60]  Abraham M Lenhoff,et al.  Three-dimensional pore structure of chromatographic adsorbents from electron tomography. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[61]  G. Carta,et al.  Protein adsorption on novel acrylamido-based polymeric ion-exchangers. IV. Effects of protein size on adsorption capacity and rate. , 2002, Journal of chromatography. A.

[62]  D. Roush,et al.  Electrostatic potentials and electrostatic interaction energies of rat cytochrome b5 and a simulated anion-exchange adsorbent surface. , 1994, Biophysical journal.

[63]  H. Brinkman A calculation of the viscous force exerted by a flowing fluid on a dense swarm of particles , 1949 .

[64]  J. A. Hubbell,et al.  Surface Treatments of Polymers for Biocompatibility , 1996 .

[65]  G. Carta,et al.  Patterns of protein adsorption in chromatographic particles visualized by optical microscopy. , 2007, Journal of chromatography. A.

[66]  E. Boschetti Advanced sorbents for preparative protein separation purposes , 1994 .

[67]  A. Staby,et al.  Comparison of chromatographic ion-exchange resins. II. More strong anion-exchange resins. , 2001, Journal of chromatography. A.

[68]  A. Seidel-Morgenstern,et al.  Fluid dynamics in monolithic adsorbents: Phenomenological approach to equivalent particle dimensions , 2002 .

[69]  N K BOARDMAN,et al.  Separation of neutral proteins on ion-exchange resins. , 1955, Nature.

[70]  Alois Jungbauer,et al.  Chromatographic media for bioseparation. , 2005, Journal of chromatography. A.

[71]  G. Carta,et al.  Protein adsorption in charged agarose gels studied by light microscopy , 2007 .

[72]  A. Lenhoff,et al.  Effects of ionic strength on lysozyme uptake rates in cation exchangers. I: Uptake in SP Sepharose FF. , 2005, Biotechnology and bioengineering.

[73]  Christian Frech,et al.  Influence of surface modification on protein retention in ion-exchange chromatography. Evaluation using different retention models. , 2009, Journal of chromatography. A.

[74]  G. Carta,et al.  Theory and applications of refractive index-based optical microscopy to measure protein mass transfer in spherical adsorbent particles. , 2008, Journal of chromatography. A.

[75]  Thomas Linden,et al.  Mechanism and kinetics of protein transport in chromatographic media studied by confocal laser scanning microscopy. Part I. The interplay of sorbent structure and fluid phase conditions. , 2003, Journal of chromatography. A.

[76]  Yan Sun,et al.  Analysis of mass transport models for protein adsorption to cation exchanger by visualization with confocal laser scanning microscopy. , 2006, Journal of chromatography. A.

[77]  Abraham M. Lenhoff,et al.  Electrostatic and van der Waals contributions to protein adsorption: computation of equilibrium constants , 1993 .

[78]  M. Plesset,et al.  Ion Exchange Kinetics. A Nonlinear Diffusion Problem , 1958 .

[79]  Giorgio Carta,et al.  Protein adsorption and transport in agarose and dextran-grafted agarose media for ion exchange chromatography. , 2007, Journal of chromatography. A.

[80]  A. G. Ogston,et al.  The spaces in a uniform random suspension of fibres , 1958 .

[81]  Christian Frech,et al.  Cation-exchange chromatography of monoclonal antibodies: Characterisation of a novel stationary phase designed for production-scale purification , 2010, mAbs.

[82]  Andreas Acrivos,et al.  Pore- and Solid-Diffusion Kinetics in Fixed-Bed Adsorption under Constant-Pattern Conditions , 1966 .

[83]  Eric J. Suda,et al.  Comparison of agarose and dextran-grafted agarose strong ion exchangers for the separation of protein aggregates. , 2009, Journal of chromatography. A.

[84]  W. Müller New ion exchangers for the chromatography of biopolymers , 1990 .

[85]  A. Lenhoff,et al.  Determination of pore size distributions of porous chromatographic adsorbents by inverse size-exclusion chromatography. , 2004, Journal of chromatography. A.

[86]  R. Denoyel,et al.  Characterisation of porous materials for bioseparation. , 2009, Journal of chromatography. A.

[87]  Peter DePhillips,et al.  Effect of spacer arm length on protein retention on a strong cation exchange adsorbent. , 2004, Analytical chemistry.

[88]  Eric J. Suda,et al.  Binding and elution behavior of proteins on strong cation exchangers. , 2009, Journal of chromatography. A.

[89]  Jörg Thömmes,et al.  Alternatives to Chromatographic Separations , 2007, Biotechnology progress.

[90]  Abhinav A Shukla,et al.  Recent advances in large-scale production of monoclonal antibodies and related proteins. , 2010, Trends in biotechnology.

[91]  I. Szleifer,et al.  Effect of molecular structure on the adsorption of protein on surfaces with grafted polymers , 2002 .

[92]  C. Horváth,et al.  Surface and pore diffusion in macroporous and gel-filled gigaporous stationary phases for protein chromatography. , 2002, Journal of chromatography. A.

[93]  G. Carta,et al.  Protein adsorption on novel acrylamido-based polymeric ion exchangers. II. Adsorption rates and column behavior. , 2000, Journal of chromatography. A.

[94]  R. Bayer,et al.  Recovery and purification process development for monoclonal antibody production , 2010, mAbs.

[95]  E. Müller Comparison between mass transfer properties of weak-anion-exchange resins with graft-functionalized polymer layers and traditional ungrafted resins. , 2003, Journal of chromatography. A.

[96]  D. Wetlaufer,et al.  Relationship between isocratic and gradient retention times in the high-performance ion-exchange chromatography of proteins. Theory and experiment. , 1986, Journal of chromatography.

[97]  Abraham M Lenhoff,et al.  Protein adsorption and transport in dextran-modified ion-exchange media. III. Effects of resin charge density and dextran content on adsorption and intraparticle uptake. , 2011, Journal of chromatography. A.

[98]  G. Carta,et al.  Protein diffusion in charged polyacrylamide gels. Visualization and analysis. , 1999, Journal of chromatography. A.

[99]  A. Lenhoff Multiscale modeling of protein uptake patterns in chromatographic particles. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[100]  A. Lenhoff,et al.  Protein adsorption and transport in dextran-modified ion-exchange media. I: adsorption. , 2009, Journal of chromatography. A.

[101]  I. Halasz,et al.  Bestimmung der Porenverteilung (10 – 4000 Å) von Festkörpern mit der Methode der Ausschluß‐Chromatographie , 1975 .

[102]  Abraham M Lenhoff,et al.  Pore size distributions of ion exchangers and relation to protein binding capacity. , 2006, Journal of chromatography. A.

[103]  C. Horváth,et al.  Combined effect of coulombic and van der Waals interactions in the chromatography of proteins. , 1992, Analytical chemistry.