Establishment and Characterisation of new Immunoreagents for diagnosis of Ovarian cancer

Abstract

Ovarian cancer is the most lethal of all gynaecological cancers and it is often diagnosed at an advanced stage due to diffuse and only mild symptoms at early stages. Early detection is crucial to increase survival but high-grade serous ovarian cancer (HGSOC) often presents non-specific symptoms, such as loss of appetite, bloating of abdomen and tiredness. This, in combination with no screening for ovarian cancer, results in over 80% of patients being diagnosed late. At this stage chemotherapy is crucial for survival and targeted therapies are often more efficient. Studies suggest that PARP inhibitors and EZH2 inhibitors are synergistic in vivo in tumours with high levels of Coactivator-associated arginine methyltransferase 1 (CARM1). Overexpression of CARM1 could therefore be a promising biomarker for precision treatment with the two substances PARP and EZH2 in combination.

This project is a collaboration between Fujirebio Diagnostics AB and an academic institute which have requested an antibody towards the biomarker CARM1. The overall goal is to establish high affinity monoclonal antibodies to the CARM1 biomarker to be used in immunohistochemistry and ultimately serum detection.

The results in this thesis indicates that creating an antibody with high specificity towards CARM1 is possible and further immunisations resulted in high serum titers in all mice. Two fusions were successfully performed and resulted in 17 hybridomas producing antibodies against CARM1. However, further work is needed to obtain antibodies with the desired properties, such as IgG isotype and high affinity against CARM1. The C-terminus GST-tagged antigen developed to screen the produced antibodies is functional and can be used for further screenings whilst cloning of the GST-tagged N-terminus antigen will need investigation and optimisation.