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Florian Capito - One of the best experts on this subject based on the ideXlab platform.
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feasibility study of semi selective Protein Precipitation with salt tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and Bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
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Feasibility study of semi‐selective Protein Precipitation with salt‐tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
Bernd Stanislawski - One of the best experts on this subject based on the ideXlab platform.
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feasibility study of semi selective Protein Precipitation with salt tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and Bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
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Feasibility study of semi‐selective Protein Precipitation with salt‐tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
Christian Schröter - One of the best experts on this subject based on the ideXlab platform.
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feasibility study of semi selective Protein Precipitation with salt tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and Bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
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Feasibility study of semi‐selective Protein Precipitation with salt‐tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
John M. Prausnitz - One of the best experts on this subject based on the ideXlab platform.
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Salt-induced Protein Precipitation: Phase equilibria from an equation of state
Fluid Phase Equilibria, 1996Co-Authors: Daniel E. Kuehner, Harvey W. Blanch, John M. PrausnitzAbstract:A molecular-thermodynamic model is developed for salt-induced Protein Precipitation. The model considers an aqueous solution of a globular Protein as a system of interacting hard spheres in a continuum pseudo-solvent (water and salt ions). The Protein molecules are considered to interact in a manner described by a set of spherically-symmetric two-body potentials of mean force. These include screened Coulombic repulsion, dispersion (van der Waals) attraction, osmotic attraction, and an attractive square-well potential intended to model specific Protein-Protein chemical interactions (including the hydrophobic effect and Protein self-associations). Following Chiew et al. (1995), an analytical equation of state is derived using the Random Phase Approximation with the hard-sphere fluid as the reference system and a perturbation based on the Protein-Protein overall potential of mean force. This equation of state provides an expression for the chemical potential of the Protein and determines liquid-liquid equilibria. The model is generalized for co-Precipitation of several Proteins. Experimental single-Protein Precipitation data are correlated for hen egg-white lysozyme and for α-chymotrypsin in concentrated aqueous solutions of ammonium sulfate.
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Molecular thermodynamics for salt-induce Protein Precipitation
AIChE Journal, 1995Co-Authors: Y.c. Chiew, Daniel E. Kuehner, Harvey W. Blanch, John M. PrausnitzAbstract:In research laboratories and in the biotechnology industry, Precipitation is commonly used to separate and isolate Proteins from solutions. A molecular-thermodynamic model is developed for salt-induced Protein Precipitation, which considers on aqueous solution of globular Protein molecules as a pseudo-one-component system containing macroions that interact through Coulombic repulsion, dispersion attraction and hydrophobic interactions, and forces arising from ion-excluded volume. Forces from ion-excluded volume take into account formation of ion pairs and ionic clusters at high salt concentrations; they are calculated in the context of the Percus-Yevick integral-equation theory. Hydrophobic interactions between exposed nonpolar amino-acid residues on the surfaces of the Protein molecules are modeled as short-range, attractive interactions between ``spherical caps`` on the surfaces of the Protein polyions. An equation of state is derived using perturbation theory. From this equation of state the authors calculate liquid-liquid equilibria: equilibrium between an aqueous phase dilute in Protein and another aqueous phase rich in Protein, which represents ``precipitated`` Protein. In the equation of state, center-to-center, spherically symmetric macroion-macroion interactions are described by the random-phase approximation, while the orientation-dependent short-range hydrophobic interaction is incorporated through the perturbation theory of associating fluids. The results suggest that either ion-excluded-volume or hydrophobic-bonding effects can precipitate Proteins inmore » aqueous solutions with high salt concentrations.« less
Harald Kolmar - One of the best experts on this subject based on the ideXlab platform.
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feasibility study of semi selective Protein Precipitation with salt tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and Bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
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Feasibility study of semi‐selective Protein Precipitation with salt‐tolerant copolymers for industrial purification of therapeutic antibodies
Biotechnology and bioengineering, 2013Co-Authors: Florian Capito, Johann Bauer, Almut Rapp, Christian Schröter, Harald Kolmar, Bernd StanislawskiAbstract:We present a feasibility study for an antibody capturing process from clarified cell culture fluid using semi-selective Protein Precipitation with salt-tolerant copolymers. Protein Precipitation is mediated by hydrophobic and electrostatic interactions with the copolymer that can be customized for the respective target. Precipitation yield with different copolymers at ionic strength of 2–22.5 mS cm−1 and pH 5.0–pH 5.7 was evaluated using pure monoclonal antibody solutions. Optimized parameters were used to elucidate yield and purity of various antibodies precipitated at physiological conditions from cell culture fluid of CHO, NS0, and SP2/0 cell culture fluid. Precipitated Protein was easily redissolved in small volume, enabling concentrating monoclonal antibodies (mAb) more than 40-fold and up to 100-fold, while residual polymer was removed to >98% using cationic polymer attached to silica flakes. mAb recovery of >90% and host cell Protein clearance of >80% were achieved, not requiring any pre-dilution of cell culture fluid. Precipitation showed no impact on mAb binding affinity when compared to non-precipitated mAb. The obtained yield and purity were lower compared to a Protein A based purification and loss of mAb was factor 1.5–3.0 higher. Yet, for high titer mAb purification processes being implemented in the future, Precipitation is an attractive option due to its ease of scalability and cost-effectiveness. Biotechnol. Bioeng. 2013;110: 2915–2927. © 2013 Wiley Periodicals, Inc.
