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Development of a Potential Recombinant Protein Vaccine in E. Coli

Recombinant proteins can be powerful stimulators of the immune system making them good candidates for vaccines. Using the intracellular parasite, Leishmania donovani as an example, this webinar will take you through the steps required for development and testing of a recombinant vaccine.

In this talk you will learn how to:

  • Express and purify protein from bacteria
  • Use the “High 3” mass spectrometry method to monitor process
  • Test the immunogenicity of the recombinant protein

Tutorial Presented By

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GenScript is the leading gene, peptide, protein and antibody research partner for fundamental life science research, translational biomedical research, and early stage pharmaceutical development. Since our establishment in 2002, GenScript has exponentially grown to become a global leading biotech company that provides life sciences services and products to scientists over 100 countries worldwide. During our tenure we have built the best-in-class capacity and capability for biological research services encompassing gene synthesis and molecular biology, peptide synthesis, custom antibodies, protein expression, antibody and protein engineering, and in vitro and in vivo pharmacology – all with the goal to Make Research Easy.

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Dr. Patrick McAtee

Tutorial Video Abstract

Expression and Purification of an Engineered, E. coli-expressed Leishmania donovani Nucleoside Hydrolase with Immunogenic Properties

Potential recombinant protein vaccine candidates must meet several criteria:

  • They must be expressed at sufficiently high levels in the organism of choice
  • They must be purified to high purity from the expression system in an immunogenic form
  • They must induce potent immune responses

Dr. Patrick McAtee will take you through these vaccine development steps using the nucleoside hydrolase antigen from Leishmania donovani as an example. He will demonstrate how his lab cloned and expressed the full-length, 36-dKa protein. He will discuss purification of the protein to >99% purity using anion exchange and gel filtration chromatography. He will also talk about the steps taken to ensure protein integrity and enzymatic activity using lithium dodecyl sulfate polyacrylamide gel electrophoresis (LDS-PAGE), mass spectrometry (MS), and enzymatic assays.

Dr. McAtee will then take you through in vivo testing of the vaccine candidate including analyzing antibody levels from mice immunized with the protein alone or in a stable emulsion with glucopyranosyl lipid adjuvant (GLA-SE). He will describe characterization of the type of cellular immune response induced by the protein. Finally, he will demonstrate protective efficacy in mice challenged with Leishmania mexicana.

E. coli Bacteria
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