E7 oncoproteins of high-risk HPV types in cervical cancer

In the present application, we propose to study the abundance of the E7 oncoproteins encoded by human papillomaviruses (HPV) of the high-risk group in cervical swabs. This technology will be used to develop new tools for the detection of tumour cells in cervical swabs, based on our observation, that the E7 oncoproteins encoded by high-risk HPVs are expressed in all cervical precancers and cancers. We propose to develop two new diagnostic strategies: on the one hand, it is proposed to use highly specific rabbit monoclonal antibodies to high- risk HPVE7 proteins for the detection of individual tumour cells in cervical smears. Conventional antibodies have been extensively tested for this purpose; however all attempts consistently failed. Our pilot studies with RabMabs indicate that the unprecedented specificity and sensitivity of these reagents will allow the detection of a few tumour cells in cervical specimens by immunohistochemistry. In a parallel approach, the same antibodies will be used to develop an ELISA method for the detection of PE7 protein in cell lysates derived from cervical smears. Based on an already available prototype ELISA that can detect picogram amounts of high-risk HPVE7 proteins, we propose to develop and clinically validate new ELISA formats with a largely increased sensitivity and specificity. Both the immunohistochemical E7 detection kit and E7 ELISA will be clinically validated with a limited number of samples during the lifetime of the project.

In the present application, we propose to study the abundance of the E7 oncoproteins encoded by human papillomaviruses (HPV) of the high-risk group in cervical swabs. This technology will be used to develop new tools for the detection of tumour cells in cervical swabs, based on our observation, that the E7 oncoproteins encoded by high-risk HPVs are expressed in all cervical precancers and cancers. We propose to develop two new diagnostic strategies: on the one hand, it is proposed to use highly specific rabbit monoclonal antibodies to high- risk HPVE7 proteins for the detection of individual tumour cells in cervical smears. Conventional antibodies have been extensively tested for this purpose; however all attempts consistently failed. Our pilot studies with RabMabs indicate that the unprecedented specificity and sensitivity of these reagents will allow the detection of a few tumour cells in cervical specimens by immunohistochemistry. In a parallel approach, the same antibodies will be used to develop an ELISA method for the detection of PE7 protein in cell lysates derived from cervical smears. Based on an already available prototype ELISA that can detect picogram amounts of high-risk HPVE7 proteins, we propose to develop and clinically validate new ELISA formats with a largely increased sensitivity and specificity. Both the immunohistochemical E7 detection kit and E7 ELISA will be clinically validated with a limited number of samples during the lifetime of the project.