New strategies for immunohistochemical analysis of biomarkers focused on rapid diagnosis of lung cancer
The aim of the project is to explore the possibility of application of innovative methodology for rapid determination of protein antigens on solid tumor cells by means of immunolabeling combined with instrumental detection beyond the sample area. One of the key elements of such diagnostic approach is the determination of PDLI and/or HER2 expression on cancer cells. As a principle, the developed method should act as support or alternative to manual assessment of tissue sections after immunohistochemical staining.
The idea of the proposed solution covers modification of immunostaining of histopathological samples (biopsies). Its key aspect lies in replacement of local staining of previously enzyme-labeled areas with a colored precipitate by an enzymatic reaction carried out in solution. As a result of the proposed modification, it would be possible to globally assess the surface density of biomarker (e.g. PDLI epitopes) in examined area. The detection process is carried out beyond the sample area and consists of instrumental readout using optical techniques (spectrophotometry or fluorometry).
Main specific goals of the research will be: i) characterization of antibody-protein antigen interactions, ii) selection of the most attractive epitopes for immunolabeling, iii) optimization of immunolabelling methodology using biopsies that have been positively verified for the presence of tumor biomarkers, iv) analysis of the usefulness of various alkaline phosphatase and peroxidase substrates from the point of view of their application in spectrophotometric or spectrofluorimetric analysis, v) studies on the correlation between classical in situ immunohistochemical staining and the proposed methodology for global biomarker determination in a biopsy sample.
Potential benefits that could arise from the proposed methodology of quantitative assessment of biomarker epitope expression in hist-pat samples include its complementarity with routine, microscopic procedures for the assessment of pathomorphological samples. Instrumental detection of biomarkers at steps of cancer qualification for treatment would open the possibility of partial replacement of subjective assessment by pathologist and/or could be an important source of information about the examined tissue. The proposed methodology, due to elimination of the subjective, microscopic assessment and its replacement with quantitative, instrumental detection of surface proteins also gives broad perspectives for miniaturization and automation of the whole process.
Taking into account the above-mentioned potential benefits, we put forward the thesis that proposed research will allow us to broaden our knowledge about cancer diagnostics and allow us to propose a new, precise and quantitative methodology for determining various epitopes of cancer cells.
As part of the BIOTECHMED-1 Research Grant “Excellence Initiative – Research University” Research Area – Biotechnology and Biomedical Engineering, WUT
Warsaw University of Technology, Centre for Advanced Materials and Technologies CEZAMAT
Warsaw University of Technology, Faculty of Chemistry
200 000 pln
Marcin Drozd, PhD Eng.