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In Silico Structural Prediction and Production of a Chimeric Recombinant Dickkopf-1 (DKK-1) Antigen

Fatemeh Malaei1, Mohammad Javad Rasaee2, Ali Mohammad Latifi3

  • 1Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. fatemeh.malaei@modares.ac.ir.

Iranian Journal of Allergy, Asthma, and Immunology
|September 16, 2019
PubMed

Insights

Researchers developed a recombinant Dickkopf-1 (rDKK-1) protein to target Wnt signaling in diseases. This rDKK-1 successfully induced a strong immune response and high antibody titers in mice.

Area of Science:

  • Biochemistry
  • Immunology
  • Molecular Biology

Background:

  • Dickkopf-1 (DKK-1) antagonizes the Wnt-β-catenin pathway.
  • DKK-1 is implicated in various cancers, including hepatocellular carcinoma and lung cancer, as well as myeloma bone disease.

Purpose of the Study:

  • To engineer a chimeric-recombinant DKK-1 (rDKK-1) protein.
  • To assess the immunogenicity of the rDKK-1 protein and its potential to induce an immune response.

Main Methods:

  • Bioinformatics analysis for designing rDKK-1.
  • Standard molecular cloning, protein expression, purification, and refolding techniques.
  • Confirmation using spectroscopy, ELISA, and Western blotting; immunogenicity tested in BALB/C mice.

Main Results:

  • Successfully designed, cloned, expressed, and purified rDKK-1 (31-kDa and 45-kDa variants).
  • Spectroscopic analysis confirmed protein structure, with CD spectra indicating stable helix structures.
  • Immunization in mice elicited a robust humoral immune response with high antibody titers.

Conclusions:

  • The engineered rDKK-1 protein is immunogenic and capable of inducing significant humoral immune responses.
  • The study demonstrates the potential of rDKK-1 as an antigen for therapeutic or diagnostic applications targeting Wnt pathway-related diseases.
Keywords:
Antibody titerChimeric antigenDickkopf-1In silico predictionRefolding

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