Establishment and Characterisation of new Immunoreagents for diagnosis of Ovarian cancer
Typ
Examensarbete för masterexamen
Master's Thesis
Master's Thesis
Program
Biotechnology (MPBIO), MSc
Publicerad
2022
Författare
Carlsson, Fanny
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
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.
Beskrivning
Ämne/nyckelord
ovarian , cancer , monoclonal , antibodies , biomarkers , CARM1 , PARPi , EZH2i