Antibody Directed Enzyme Prodrug Therapy (ADEPT) is a technique used in cancer treatment, which convert a prodrug to a powerful cytotoxic drug only in the vicinity of the tumor. The technique relies on a bacterial enzyme, glucarpidase (former name: carboxypeptidase G2, CPG2). Also the glucarpidase is a very effective enzyme for detoxification of methotrexate, (MTX) which serves as an important component of various chemotherapeutic regimens for the treatment of cancer patients.

Repeated cycles of ADEPT and the use of wild type glucarpidase in detoxification are essential but are hampered by the human antibody response to the enzyme. Additionally, glucarpidase has a relatively slow action in detoxification.

The aim of the work is to provide solutions to overcome the pitfalls of the techniques through the application of different strategies.

In our work we successfully isolated new glucarpidase produces from soils. We cloned and overexpressed the novel gene in E.coli. The new recombinant glucarpidase has been characterized. We also produce different variants of our new gene using different strategies. These variants will be investigated to isolate glucarpidase with higher activity and glucarpidase which avoid the immune system.


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