A critical goal in prevention and control of cancer is early detection of cancer. it is suggested that the dickkopf1 (dkk1) has the ability to act as tumor marker in early detection. the aim of the present study was to produce rdkk1.
Methods
A synthetic chimeric gene, encoding comp, p2 and p30 epitopes from tetanus toxin and truncated parts of the dkk1 was designed and optimized based on bioinformatics studies. chimeric gene was cloned, expressed, purified and then its folding was evaluated.
Results
The stronger immunogenic parts were selected and the solubility of chimeric dkk1 antigen was improved by bioinformatics’ evaluation.
pcr, sds-page (sodium dodecyl sulfate polyacrylamide gel electrophoresis), and enzyme-linked immunosorbent assay (elisa) were used to confirm rdkk1 cloning, expression, purification and refolding. the designed chimeric protein showed a good elisa response to the commercial polyclonal dkk1 antibody.
Conclusion
Results show that rdkk1 can be used as an antigen to produce antibody against dkk1 protein.
Keywords
Dickkopf-1, antigen, chimeric protein, protein refolding, protein purification
