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Related Experiment Videos

Selection and characterization of Her2 binding-designed ankyrin repeat proteins.

Christian Zahnd1, Frédéric Pecorari, Nadine Straumann

  • 1Department of Biochemistry, University of Zürich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland.

The Journal of Biological Chemistry
|September 12, 2006
PubMed
Summary

Designed ankyrin repeat proteins (DARPins) were developed against human epidermal growth factor receptor 2 (Her2). These DARPins show high affinity and specificity, suggesting potential for cancer diagnostics and therapy.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • Designed ankyrin repeat proteins (DARPins) are engineered binding proteins with high affinity and specificity.
  • Human epidermal growth factor receptor 2 (Her2) is a key target in cancer therapy and diagnosis.
  • Existing therapies like trastuzumab and pertuzumab target Her2 but have specific epitope binding.

Purpose of the Study:

  • To select and characterize DARPins that bind to Her2.
  • To evaluate the binding affinity, specificity, and diagnostic potential of selected DARPins.
  • To explore the utility of DARPins in cancer diagnostics and targeted therapy.

Main Methods:

  • In vitro selection of DARPins against Her2.
  • Affinity determination using Kd measurements.

Related Experiment Videos

  • Specificity testing against Her2, EGFR1, and other proteins.
  • Cell line binding assays (SKBR-3, MCF-7).
  • Immunohistochemical staining of human breast carcinoma tissue.

    • Main Results:

    • Several DARPins were selected, with some binding to the same epitope as trastuzumab.
    • Selected DARPins exhibited low nanomolar binding affinity (Kd=7.3 nm).
    • DARPins demonstrated high specificity, with no cross-reactivity to EGFR1 or other tested proteins.
    • DARPins effectively recognized Her2 on both high- and low-expressing cancer cell lines.
    • Specific staining of Her2 on human breast carcinoma tissue sections was observed.

    Conclusions:

    • Selected DARPins are highly specific and bind Her2 with high affinity.
    • These DARPins show promise for developing diagnostic tests for Her2 overexpression in adenocarcinomas.
    • The favorable properties of DARPins make them suitable candidates for further evaluation in targeted cancer therapy, including fusion protein development.