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J. Hu

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    Poster Session (ID 8)

    • Event: ACLC 2018
    • Type: Poster Session
    • Track:
    • Presentations: 1
    • Moderators:
    • Coordinates: 11/07/2018, 00:00 - 00:00, Poster Hall
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      P033 - A Fusion-Protein Approach Enabling Mammalian Cell Production of Tumor Targeting Protein Domains for Therapeutic Development (ID 171)

      00:00 - 00:00  |  Author(s): J. Hu

      • Abstract

      Background: Single chain Fv fragment (scFv) is kind of fusion protein of the variable regions of heavy (VH) and light chains (VL) of immunoglobulins, and it is an important element of chimeric antigen receptors (CARs) for cancer therapy.

      Methods: We sought to produce a panel of 16 extracellular protein domains of tumor markers for use in scFv yeast library screenings. We developed a series of vectors comprising various combinations of expression elements and made GC content optimization of the signal peptide.

      Results: Overall, expression was unpredictable and more than half of the protein domains could not be produced using any of the constructs. Here we described a novel fusion expression system based on mouse TEM7 (tumor endothelial marker 7), which could facilitate protein expression. With this approach we could produce all but one of the tumor marker domains that could not otherwise be expressed. In addition, we demonstrated that the tumor associated antigen hFZD10 produced as a fusion protein with mTEM7 could be used to enrich scFv antibodies from a yeast display library.

      Conclusion: Collectively our study demonstrates the potential of specific fusion proteins enabling mammalian cell production of tumor targeting protein domains for therapeutic development.