Optimization of downstream processing for plasmid DNA using ion-exchange chromatography

POSTER

Abstract

Increasing interest in gene therapy and DNA vaccines has sparked an industry-wide focus on producing large quantities of plasmid DNA, while still maintaining quality requirements from regulatory agencies. Optimization of the downstream processing of pDNA has shown promising results for high quality, high quantity batches of plasmids. A protocol was developed and optimized for ion-exchange capture and purification of 5kpb pDNA extracted from E. Coli lysate using commercial borosilicate membrane filters. When compared to similar published processes using varying mediums, our protocol was as effective or more effective in maintaining quality, purity and yield. Principles and procedures developed in this protocol will help aid further research in adapting this process to a newly developed filter medium that offers extremely high surface area for increased binding capacity.

Authors

  • Camille Baird

    Brigham Young University - Provo

  • William Stockwell

    Logan High School, University of Tsukuba, Department of Physics, University of Texas at El Paso, El Paso, TX, Division of Engineering, Mayo Clinic, Rochester, MN, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Utah Valley University, University of New Mexico, Brigham Young University, Moxtek, INC, New Mexico State University, Oak Ridge National Laboratory, Oak Ridge TN 37830, USA, Los Alamos National Laboratory, Argonne National Laboratory, Embry-Riddle Aeronautical University, University of Newcastle, University of Sydney, Brigham Young University Provo, Rice University, Perimeter Institute and University of Guelph, University of Reims Champagne-Ardenne, France, Texas A&M University, University of Illinois at Urbana-Champaign, University of Valencia, Spain, None, University of Texas at El Paso, Universidad Simon Bolivar, Universidad Autonoma de Ciudad Juarez, Brigham Young University - Provo, Morgan High School - Morgan, UT, Utah State University, University of Tsukaba, Chemistry Research Center, US Air Force Academy, USAFA, JILA, University of Colorado Boulder, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX, Department of Orthopedics, Mayo Clinic, Rochester, MN, Colorado State University, National Research Tomsk Polytechnic University \& New Mexico State University,, National Institute of Standards and Technology Center for Neutron Research, Oak Ridge National Laboratory, Huazhong University of Science and Technology, U of Utah, NMSU, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Chemical Science Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA, INM - Leibniz Institute for New Materials, Saarbrucken, Germany, Department of Physics, The University of Texas at El Paso, El Paso, Texas, 7996, UTEP, Arizona State University, The University of Texas at El Paso, none, National Renewable Energy Laboratory, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, Division of Engineering, Mayo Clinic, Department of Neurologic Surgery, Mayo Clinic, Department of Physics & Astronomy, Texas A&M University, Commerce,TX-75428, USA., INFN, Sezione di Pisa, Largo Pontecorvo 3, 56127 Pisa, Italy, Texas A&M University Commerce, Commerce, Texas-75429, Department of Physics and Astronomy, Texas A&M University-Commerce, Commerce, Texas 75429, USA, Cyclotron Institute, Texas A&M University, College Station, Texas 77843, USA, Department of Physics, University of Texas at El Paso, University of Delaware, IIT INDORE, UT AUSTIN, Idaho Accelerator Center, Brigham Young University, Physics and Astronomy, Univesidad de Guanajuato, Fermilab, École Polytechnique Fédérale de Lausanne, RadiaSoft, LLC, Weber State University, University of Texas at Dallas, National High Magnetic Field Laboratory, University of Texas Southwestern Medical Center, University of Alabama in Huntsville, Marshall Space Flight Center, NASA, Howard University

  • William Stockwell

    Logan High School, University of Tsukuba, Department of Physics, University of Texas at El Paso, El Paso, TX, Division of Engineering, Mayo Clinic, Rochester, MN, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Utah Valley University, University of New Mexico, Brigham Young University, Moxtek, INC, New Mexico State University, Oak Ridge National Laboratory, Oak Ridge TN 37830, USA, Los Alamos National Laboratory, Argonne National Laboratory, Embry-Riddle Aeronautical University, University of Newcastle, University of Sydney, Brigham Young University Provo, Rice University, Perimeter Institute and University of Guelph, University of Reims Champagne-Ardenne, France, Texas A&M University, University of Illinois at Urbana-Champaign, University of Valencia, Spain, None, University of Texas at El Paso, Universidad Simon Bolivar, Universidad Autonoma de Ciudad Juarez, Brigham Young University - Provo, Morgan High School - Morgan, UT, Utah State University, University of Tsukaba, Chemistry Research Center, US Air Force Academy, USAFA, JILA, University of Colorado Boulder, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX, Department of Orthopedics, Mayo Clinic, Rochester, MN, Colorado State University, National Research Tomsk Polytechnic University \& New Mexico State University,, National Institute of Standards and Technology Center for Neutron Research, Oak Ridge National Laboratory, Huazhong University of Science and Technology, U of Utah, NMSU, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Chemical Science Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA, INM - Leibniz Institute for New Materials, Saarbrucken, Germany, Department of Physics, The University of Texas at El Paso, El Paso, Texas, 7996, UTEP, Arizona State University, The University of Texas at El Paso, none, National Renewable Energy Laboratory, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, Division of Engineering, Mayo Clinic, Department of Neurologic Surgery, Mayo Clinic, Department of Physics & Astronomy, Texas A&M University, Commerce,TX-75428, USA., INFN, Sezione di Pisa, Largo Pontecorvo 3, 56127 Pisa, Italy, Texas A&M University Commerce, Commerce, Texas-75429, Department of Physics and Astronomy, Texas A&M University-Commerce, Commerce, Texas 75429, USA, Cyclotron Institute, Texas A&M University, College Station, Texas 77843, USA, Department of Physics, University of Texas at El Paso, University of Delaware, IIT INDORE, UT AUSTIN, Idaho Accelerator Center, Brigham Young University, Physics and Astronomy, Univesidad de Guanajuato, Fermilab, École Polytechnique Fédérale de Lausanne, RadiaSoft, LLC, Weber State University, University of Texas at Dallas, National High Magnetic Field Laboratory, University of Texas Southwestern Medical Center, University of Alabama in Huntsville, Marshall Space Flight Center, NASA, Howard University

  • William Stockwell

    Logan High School, University of Tsukuba, Department of Physics, University of Texas at El Paso, El Paso, TX, Division of Engineering, Mayo Clinic, Rochester, MN, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Utah Valley University, University of New Mexico, Brigham Young University, Moxtek, INC, New Mexico State University, Oak Ridge National Laboratory, Oak Ridge TN 37830, USA, Los Alamos National Laboratory, Argonne National Laboratory, Embry-Riddle Aeronautical University, University of Newcastle, University of Sydney, Brigham Young University Provo, Rice University, Perimeter Institute and University of Guelph, University of Reims Champagne-Ardenne, France, Texas A&M University, University of Illinois at Urbana-Champaign, University of Valencia, Spain, None, University of Texas at El Paso, Universidad Simon Bolivar, Universidad Autonoma de Ciudad Juarez, Brigham Young University - Provo, Morgan High School - Morgan, UT, Utah State University, University of Tsukaba, Chemistry Research Center, US Air Force Academy, USAFA, JILA, University of Colorado Boulder, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX, Department of Orthopedics, Mayo Clinic, Rochester, MN, Colorado State University, National Research Tomsk Polytechnic University \& New Mexico State University,, National Institute of Standards and Technology Center for Neutron Research, Oak Ridge National Laboratory, Huazhong University of Science and Technology, U of Utah, NMSU, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Chemical Science Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA, INM - Leibniz Institute for New Materials, Saarbrucken, Germany, Department of Physics, The University of Texas at El Paso, El Paso, Texas, 7996, UTEP, Arizona State University, The University of Texas at El Paso, none, National Renewable Energy Laboratory, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, Division of Engineering, Mayo Clinic, Department of Neurologic Surgery, Mayo Clinic, Department of Physics & Astronomy, Texas A&M University, Commerce,TX-75428, USA., INFN, Sezione di Pisa, Largo Pontecorvo 3, 56127 Pisa, Italy, Texas A&M University Commerce, Commerce, Texas-75429, Department of Physics and Astronomy, Texas A&M University-Commerce, Commerce, Texas 75429, USA, Cyclotron Institute, Texas A&M University, College Station, Texas 77843, USA, Department of Physics, University of Texas at El Paso, University of Delaware, IIT INDORE, UT AUSTIN, Idaho Accelerator Center, Brigham Young University, Physics and Astronomy, Univesidad de Guanajuato, Fermilab, École Polytechnique Fédérale de Lausanne, RadiaSoft, LLC, Weber State University, University of Texas at Dallas, National High Magnetic Field Laboratory, University of Texas Southwestern Medical Center, University of Alabama in Huntsville, Marshall Space Flight Center, NASA, Howard University

  • William Stockwell

    Logan High School, University of Tsukuba, Department of Physics, University of Texas at El Paso, El Paso, TX, Division of Engineering, Mayo Clinic, Rochester, MN, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Utah Valley University, University of New Mexico, Brigham Young University, Moxtek, INC, New Mexico State University, Oak Ridge National Laboratory, Oak Ridge TN 37830, USA, Los Alamos National Laboratory, Argonne National Laboratory, Embry-Riddle Aeronautical University, University of Newcastle, University of Sydney, Brigham Young University Provo, Rice University, Perimeter Institute and University of Guelph, University of Reims Champagne-Ardenne, France, Texas A&M University, University of Illinois at Urbana-Champaign, University of Valencia, Spain, None, University of Texas at El Paso, Universidad Simon Bolivar, Universidad Autonoma de Ciudad Juarez, Brigham Young University - Provo, Morgan High School - Morgan, UT, Utah State University, University of Tsukaba, Chemistry Research Center, US Air Force Academy, USAFA, JILA, University of Colorado Boulder, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX, Department of Orthopedics, Mayo Clinic, Rochester, MN, Colorado State University, National Research Tomsk Polytechnic University \& New Mexico State University,, National Institute of Standards and Technology Center for Neutron Research, Oak Ridge National Laboratory, Huazhong University of Science and Technology, U of Utah, NMSU, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Chemical Science Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA, INM - Leibniz Institute for New Materials, Saarbrucken, Germany, Department of Physics, The University of Texas at El Paso, El Paso, Texas, 7996, UTEP, Arizona State University, The University of Texas at El Paso, none, National Renewable Energy Laboratory, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, Division of Engineering, Mayo Clinic, Department of Neurologic Surgery, Mayo Clinic, Department of Physics & Astronomy, Texas A&M University, Commerce,TX-75428, USA., INFN, Sezione di Pisa, Largo Pontecorvo 3, 56127 Pisa, Italy, Texas A&M University Commerce, Commerce, Texas-75429, Department of Physics and Astronomy, Texas A&M University-Commerce, Commerce, Texas 75429, USA, Cyclotron Institute, Texas A&M University, College Station, Texas 77843, USA, Department of Physics, University of Texas at El Paso, University of Delaware, IIT INDORE, UT AUSTIN, Idaho Accelerator Center, Brigham Young University, Physics and Astronomy, Univesidad de Guanajuato, Fermilab, École Polytechnique Fédérale de Lausanne, RadiaSoft, LLC, Weber State University, University of Texas at Dallas, National High Magnetic Field Laboratory, University of Texas Southwestern Medical Center, University of Alabama in Huntsville, Marshall Space Flight Center, NASA, Howard University

  • William Stockwell

    Logan High School, University of Tsukuba, Department of Physics, University of Texas at El Paso, El Paso, TX, Division of Engineering, Mayo Clinic, Rochester, MN, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Utah Valley University, University of New Mexico, Brigham Young University, Moxtek, INC, New Mexico State University, Oak Ridge National Laboratory, Oak Ridge TN 37830, USA, Los Alamos National Laboratory, Argonne National Laboratory, Embry-Riddle Aeronautical University, University of Newcastle, University of Sydney, Brigham Young University Provo, Rice University, Perimeter Institute and University of Guelph, University of Reims Champagne-Ardenne, France, Texas A&M University, University of Illinois at Urbana-Champaign, University of Valencia, Spain, None, University of Texas at El Paso, Universidad Simon Bolivar, Universidad Autonoma de Ciudad Juarez, Brigham Young University - Provo, Morgan High School - Morgan, UT, Utah State University, University of Tsukaba, Chemistry Research Center, US Air Force Academy, USAFA, JILA, University of Colorado Boulder, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX, Department of Orthopedics, Mayo Clinic, Rochester, MN, Colorado State University, National Research Tomsk Polytechnic University \& New Mexico State University,, National Institute of Standards and Technology Center for Neutron Research, Oak Ridge National Laboratory, Huazhong University of Science and Technology, U of Utah, NMSU, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Rochester, MN, Chemical Science Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA, INM - Leibniz Institute for New Materials, Saarbrucken, Germany, Department of Physics, The University of Texas at El Paso, El Paso, Texas, 7996, UTEP, Arizona State University, The University of Texas at El Paso, none, National Renewable Energy Laboratory, Division of Engineering, Department of Neurologic Surgery, Mayo Clinic, Department of Physics, Border Biomedical Research Center, University of Texas at El Paso, Division of Engineering, Mayo Clinic, Department of Neurologic Surgery, Mayo Clinic, Department of Physics & Astronomy, Texas A&M University, Commerce,TX-75428, USA., INFN, Sezione di Pisa, Largo Pontecorvo 3, 56127 Pisa, Italy, Texas A&M University Commerce, Commerce, Texas-75429, Department of Physics and Astronomy, Texas A&M University-Commerce, Commerce, Texas 75429, USA, Cyclotron Institute, Texas A&M University, College Station, Texas 77843, USA, Department of Physics, University of Texas at El Paso, University of Delaware, IIT INDORE, UT AUSTIN, Idaho Accelerator Center, Brigham Young University, Physics and Astronomy, Univesidad de Guanajuato, Fermilab, École Polytechnique Fédérale de Lausanne, RadiaSoft, LLC, Weber State University, University of Texas at Dallas, National High Magnetic Field Laboratory, University of Texas Southwestern Medical Center, University of Alabama in Huntsville, Marshall Space Flight Center, NASA, Howard University