Impact of Ligand Density on the Optimization of Ion-Exchange Membrane Chromatography for Viral Vector Purification

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Abstract

The effect of ligand density on anion-exchange membrane chromatography (AEXmc) for the purification of recombinant baculoviruses (rBVs), potential viral vectors in clinical applications, is studied by surface plasmon resonance on customized AEX surfaces and gradient elution experiments on Sartobind D membrane prototypes with different diethylamine ligand densities, complemented by dynamic light scattering analysis for estimation of the hydrodynamic particle size of the various biologics. A chromatographic-column model based on the steric mass action model of ion exchange is employed to analyze the gradient-elution AEXmc experiments, extrapolate the results to other operating conditions, and provide directions for process improvement. Although counterintuitively, the experimental evidence provided in this study shows that the lowering of ligand density is beneficial for rBV purification by AEXmc in bind-and-elute-mode, because it decreases the residual concentrations of host cell protein, dsDNA, and non-infective rBVs in the eluted product cut, and increases the overall yield by roughly 20% over current standard values. Overall, we present a case study on how rational design can streamline downstream process development. Biotechnol. Bioeng. 2011; 108: 1347-1359. (C) 2011 Wiley Periodicals, Inc.
Original languageUnknown
Pages (from-to)1347-1359
JournalBiotechnology and Bioengineering
Volume108
Issue number6
DOIs
Publication statusPublished - 1 Jan 2011

Keywords

    Cite this

    @article{02f6d6f4edfc4698a5e00843a73298e0,
    title = "Impact of Ligand Density on the Optimization of Ion-Exchange Membrane Chromatography for Viral Vector Purification",
    abstract = "The effect of ligand density on anion-exchange membrane chromatography (AEXmc) for the purification of recombinant baculoviruses (rBVs), potential viral vectors in clinical applications, is studied by surface plasmon resonance on customized AEX surfaces and gradient elution experiments on Sartobind D membrane prototypes with different diethylamine ligand densities, complemented by dynamic light scattering analysis for estimation of the hydrodynamic particle size of the various biologics. A chromatographic-column model based on the steric mass action model of ion exchange is employed to analyze the gradient-elution AEXmc experiments, extrapolate the results to other operating conditions, and provide directions for process improvement. Although counterintuitively, the experimental evidence provided in this study shows that the lowering of ligand density is beneficial for rBV purification by AEXmc in bind-and-elute-mode, because it decreases the residual concentrations of host cell protein, dsDNA, and non-infective rBVs in the eluted product cut, and increases the overall yield by roughly 20{\%} over current standard values. Overall, we present a case study on how rational design can streamline downstream process development. Biotechnol. Bioeng. 2011; 108: 1347-1359. (C) 2011 Wiley Periodicals, Inc.",
    keywords = "RETROVIRAL VECTORS, VIRUSES, ADSORPTION, PROTEIN, ADENOVIRUS, CELLS, SURFACE-PLASMON RESONANCE, SEPARATIONS, BACULOVIRUS",
    author = "Mota, {Jos{\'e} Paulo Barbosa} and Carrondo, {Manuel Jos{\'e} Teixeira} and Alves, {Paula Maria}",
    year = "2011",
    month = "1",
    day = "1",
    doi = "10.1002/bit.23058",
    language = "Unknown",
    volume = "108",
    pages = "1347--1359",
    journal = "Biotechnology and Bioengineering",
    issn = "0006-3592",
    publisher = "WILEY-BLACKWELL",
    number = "6",

    }

    TY - JOUR

    T1 - Impact of Ligand Density on the Optimization of Ion-Exchange Membrane Chromatography for Viral Vector Purification

    AU - Mota, José Paulo Barbosa

    AU - Carrondo, Manuel José Teixeira

    AU - Alves, Paula Maria

    PY - 2011/1/1

    Y1 - 2011/1/1

    N2 - The effect of ligand density on anion-exchange membrane chromatography (AEXmc) for the purification of recombinant baculoviruses (rBVs), potential viral vectors in clinical applications, is studied by surface plasmon resonance on customized AEX surfaces and gradient elution experiments on Sartobind D membrane prototypes with different diethylamine ligand densities, complemented by dynamic light scattering analysis for estimation of the hydrodynamic particle size of the various biologics. A chromatographic-column model based on the steric mass action model of ion exchange is employed to analyze the gradient-elution AEXmc experiments, extrapolate the results to other operating conditions, and provide directions for process improvement. Although counterintuitively, the experimental evidence provided in this study shows that the lowering of ligand density is beneficial for rBV purification by AEXmc in bind-and-elute-mode, because it decreases the residual concentrations of host cell protein, dsDNA, and non-infective rBVs in the eluted product cut, and increases the overall yield by roughly 20% over current standard values. Overall, we present a case study on how rational design can streamline downstream process development. Biotechnol. Bioeng. 2011; 108: 1347-1359. (C) 2011 Wiley Periodicals, Inc.

    AB - The effect of ligand density on anion-exchange membrane chromatography (AEXmc) for the purification of recombinant baculoviruses (rBVs), potential viral vectors in clinical applications, is studied by surface plasmon resonance on customized AEX surfaces and gradient elution experiments on Sartobind D membrane prototypes with different diethylamine ligand densities, complemented by dynamic light scattering analysis for estimation of the hydrodynamic particle size of the various biologics. A chromatographic-column model based on the steric mass action model of ion exchange is employed to analyze the gradient-elution AEXmc experiments, extrapolate the results to other operating conditions, and provide directions for process improvement. Although counterintuitively, the experimental evidence provided in this study shows that the lowering of ligand density is beneficial for rBV purification by AEXmc in bind-and-elute-mode, because it decreases the residual concentrations of host cell protein, dsDNA, and non-infective rBVs in the eluted product cut, and increases the overall yield by roughly 20% over current standard values. Overall, we present a case study on how rational design can streamline downstream process development. Biotechnol. Bioeng. 2011; 108: 1347-1359. (C) 2011 Wiley Periodicals, Inc.

    KW - RETROVIRAL VECTORS

    KW - VIRUSES

    KW - ADSORPTION

    KW - PROTEIN

    KW - ADENOVIRUS

    KW - CELLS

    KW - SURFACE-PLASMON RESONANCE

    KW - SEPARATIONS

    KW - BACULOVIRUS

    U2 - 10.1002/bit.23058

    DO - 10.1002/bit.23058

    M3 - Article

    VL - 108

    SP - 1347

    EP - 1359

    JO - Biotechnology and Bioengineering

    JF - Biotechnology and Bioengineering

    SN - 0006-3592

    IS - 6

    ER -