DEC 11, 2019 07:00 AM PST
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A Sensitive and Robust Plasmid Analysis Method by Capillary Electrophoresis with Laser-Induced Fluorescence Detection

SPONSORED BY: SCIEX
C.E. CREDITS: P.A.C.E. CE | Florida CE
Speakers
  • Scientist in Analytical Research and Development, Pfizer
    Biography
      Ms. Reed received her Bachelor's Degree in Cell and Molecular Biology from Missouri State University. After graduating from college, she joined Wyeth Biopharma in St Louis, working on the Refacto FVIII therapy and gaining ample experiences in QC analyticals.

      After that, she joined the biologics formulation development group at Pfizer in St. Louis. A few years later she transitioned to biologics Analytical Research and Development line, working with a diverse group of analytical technologies, and eventually adopting CE as her main technology focus.

      She has worked with the Sciex CE technology for about 11 years by now, most recently moving into a role of method development for novel biological projects. Her current work is focused on development of methods to evaluate critical quality attributes to be validated in support of clinical trials for gene therapy projects and BLA submissions.

    Abstract:
    DATE:   December 11, 2019
    TIME:    7:00am PST, 10:00am EST
     
    An important part of the biopharma industry, Plasmid DNA is used as a gene-delivery vehicle for DNA vaccination and as a key intermediate for processes like viral particle production for gene therapy and ex-vivo protein synthesis. Most plasmid DNA preparations contain several topological variants or isoforms including the supercoiled, open-circular and linear forms of the molecule.
     
    Federal regulations require purity testing for manufactured injectable plasmid products and recommend establishing a release criterion of > 80% supercoiled content. Unfortunately, traditional agarose gel has limitations on quantitation. Capillary electrophoresis with Laser Induced Fluorescence detection (CE-LIF) provides a rapid, sensitive, reproducible and automated method for the quantitative analysis of plasmid DNA isoforms.
     
    In this webinar, we present a method for plasmid analysis by CE-LIF with the use of a common dye. Results obtained with two large plasmids at 7 to 10 kb demonstrate that this method is sensitive and robust, providing baseline resolution of supercoiled, open-circular and linear plasmid isoforms within 20 minutes, and is suitable for testing plasmid purity and monitoring plasmid degradation.
     
    Learning Objectives:
    • Demonstrate fast analysis of plasmid samples with separation time within 12 minutes
    • Identify Baseline resolution of supercoiled, linear and open circular isoforms of a 5 kb and a 7 kb plasmid
    • Illustrate streamlining workflows using commercially available dsDNA 1000 kit and LIFluor EnhanCE stain
    • Explain automated sample separation and data analysis provide rapid, reproducible and quantitative results with RSD of 0.23% for migration time and 0.61% for % area 

     

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    LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.

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