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Opn final 10 19-10

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Development of monoclonal antibodies to Osteopontin

Development of monoclonal antibodies to Osteopontin

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  • Additional studies currently underway include a western blot analysis using native purified OPN against all six of the selected clones as well as cell adhesion assays using the native OPN. If the DSVVYG peptide is linear, the western blot data will shed light on the integrity of the purified native OPN by immunostaining full length and /or OPN fragments containing the DSVVYG, indicated by molecular weight of bands. Additionally, the ability of these clones to inhibit cell adhesion by binding to native OPN will be of significant value to cancer researchers and is the motivation to continue these studies. Results of the extended studies will be made available at a later date. Please contact Sales to access samples of this unique panel of Osteopontin Monoclonal antibodies.

Opn final 10 19-10 Presentation Transcript

  • 1. Monoclonal Antibodies Specific for DSVVYG Peptide of Human Osteopontin
  • 2. Functional Domain Structure of Osteopontin
    Immunogen to target cryptic site of OPN: DSVVYG
  • 3. Assays: Indirect ELISA
    Peptide
    Recombinant N-term OPN
    Recombinant Full Length OPN
    Native OPN (purified from human breast milk), evaluated on SDS-PAGE
  • 4. Clones Selected for Further Study
  • 5. Assays: Sandwich ELISA
    Nine selected peptide clones as tracer with current 5 MABS as capture, rec-flOPNand native OPN as antigen
    Nine selected peptide clones as capture with current 5 MABS as tracer, rec-flOPNand native OPN as antigen
  • 6. Matched Pair ELISA, Peptide clones Tracer
    Native OPN
    flOPN
  • 7. Matched Pair ELISA, Peptide clones Capture
    Native OPN
    flOPN
  • 8. Cell Adhesion AssaysBreast Cancer Cell Line: MDA-MB-231
  • 9. Conclusions
    MBS has developed a panel of antibodies specific to the unique DSVVYG sequence of OPN
    Six clones emerged based on the data generated from ELISA and Cell Adhesion as being of most interest.
    Data demonstrates that using native OPN in the assays is optimal, as recombinantly expressed OPN in E. coli will not provide the post translational modifications of native OPN
    Samples of the six clones are available upon request
  • 10. Next Actions
    Study of Peptide specific OPN clones by Western Blot of Native OPN
    Study of Peptide Specific OPN clones on Cell Adhesion Assays using Native OPN
    References: Alicia Plumer, HongyiDuan, SripriyaSubramaniam, F Lee Lucas, Susan Miesfeldt, Ah-Kau Ng, Lucy Liaw. Development of fragment specific osteopontin antibodies and ELISA for quantification in human metastatic breast cancer BMC Cancer 2008, 8:38 (31 January 2008)