The objectives of the Phase II project are to further develop a new protease detection technology platform with versatile application capabilities (in vitro and in vivo) including cancer applications: detection of two markers within a common signaling pathway to determine the prognosis of ovarian and breast cancer. The Phase II project will complete the analytical characterization of the assays as described in """"""""Bioanalytical Methods Validation: Guidance for Industry"""""""", US DHHS, FDA, COER, CVV, May 2001.
Aim 1 begins the process by developing and characterizing assay performance using purified proteases in simple reconstituted samples.
Aim 2 continues the move towards clinical sample by examining sample processing, stability, and matrix effects through the use of commercially available normal tissue and tumor tissues. Finally, Aims 3 and 4 describe the characterization of protease levels across a variety of tissue biopsy samples for breast and ovarian cancers to provide an indication of the range of protease activity levels across samples of different types and patient to patient variability within each tissue type. These steps are necessary precursors to any subsequent development and validation through clinical trials of the protease assays as prognostic tools