Assay development
Multi-omics Biomarker Pipeline Reveals Elevated Levels of Protein-glutamine Gamma-glutamyltransferase 4 in Seminal Plasma of Prostate Cancer PatientsMulti-omics biomarker pipeline generated 19 proteins which were verified by a multiplex quantitative SRM assay in seminal plasma samples of 152 prostate cancer and 67 negative biopsy patients. Verification revealed a prostate-specific, secreted and androgen-regulated protein-glutamine gamma-glutamyltransferase 4 (TGM4), which detected prostate cancer on biopsies with AUC=0.66, as measured by an in-house immunoassay in seminal plasma. Perspectives of seminal plasma proteins as biomarkers of prostate cancer were reviewed.
Intact Transition Epitope Mapping – Targeted High-Energy Rupture of Extracted Epitopes (ITEM-THREE)ITEM-THREE enables rapid epitope mapping. Sample consumption is minimized and in-solution handling reduced to mixing of antibody and antigen peptide solutions. After immune complex formation in solution, epitope mapping is performed in the gas phase using the mass spectrometer for sophisticated ion manipulation and filtering. Because amino acid sequence information is obtained from the epitope peptide, unknown antigens can be identified. Knowing the epitope broadens the application of antibodies to unspecified target proteins from any organism.
High-throughput Identification of FLT3 Wild-type and Mutant Kinase Substrate Preferences and Application to Design of Sensitive In Vitro Kinase Assay SubstratesKinase activity assays are an invaluable tool used to develop new small molecule inhibitors to combat cancer and other human diseases, however, only a small fraction of the human kinome is well characterized. Lack of biological substrate information limits the capability of computational approaches to design sensitive peptide probes for FLT3 and clinically relevant mutant kinases. In this report, we used phosphoproteomics and the KINATEST-ID pipeline (31) to design a panel of FLT3 artificial peptide substrates (FAStides) to design an ELISA-based activity assay.