ABSTRACT
ABSTRACT Early-stage detection of cancer is the key to provide a better outcome for therapeutic intervention. Most routine screening tools for cancer detection are largely based on examination of cell morphology, tissue histology, and measurement of serum markers, which lack sufcient sensitivity and/or specicity for early detection of cancer. Indeed, most secreted proteins studied as cancer screening biomarkers have low sensitivity and/or low specicity, and this could be due to the use of nonsensitive techniques or due the fact that several of these tumor markers are also produced by normal tissues. Altogether, these facts pinpoint to the urgent need for the discovery of innovative tools and novel tumor
4.1 Real-Time PCR for the Detection of Wilms’ Tumor Gene 1 in Cancer ........................................ 48 4.1.1 Introduction ....................................................................................................................... 48 4.1.2 WT1 Isoforms .................................................................................................................... 49 4.1.3 Experimental Procedures .................................................................................................. 50
4.1.3.1 Sample Collection, Processing, and RNA Isolation .......................................... 50 4.1.3.2 Reverse Transcription PCR for WT1 ................................................................. 50 4.1.3.3 Real-Time qPCR for WT1 ..................................................................................51 4.1.3.4 Result Interpretation of the qPCR ..................................................................... 54
4.1.4 Challenges and Future Directions ..................................................................................... 54 4.2 Mass Spectrometry-Based Proteomics for Detection of Cancer ................................................... 55
4.2.1 Introduction ....................................................................................................................... 55 4.2.2 Proteomic Approaches for Cancer Studies ....................................................................... 55
4.2.2.1 Biomarker Discovery ......................................................................................... 58 4.2.2.2 Clinical Diagnostics ........................................................................................... 60
4.2.3 Proteomic Platforms for Cancer Studies ........................................................................... 60 4.2.3.1 SELDI-TOF ........................................................................................................ 60 4.2.3.2 MALDI-TOF .......................................................................................................61 4.2.3.3 1D and 2D Gels/MS ........................................................................................... 62 4.2.3.4 LC-ESI-MS (/MS) .............................................................................................. 63
4.2.4 Challenges and Future Directions ..................................................................................... 64 4.2.4.1 Limitations and Challenges ............................................................................... 64 4.2.4.2 Future Directions ............................................................................................... 66
4.3 Surface Plasmon Resonance for Detection of Cancer ................................................................... 67 4.3.1 Introduction ....................................................................................................................... 67 4.3.2 SPR Principles ................................................................................................................... 67 4.3.3 Application of SPR in Cancer Biomarker Discovery ........................................................ 69 4.3.4 Integration of SPR with Mass Spectrometry .................................................................... 70 4.3.5 Challenges and Future Directions ..................................................................................... 73
4.4 Considerations for Biological Samples .......................................................................................... 73 References ................................................................................................................................................ 74
markers for cancer screening, diagnosis, and prognosis. Recently, scientists and clinicians have shifted to innovative techniques in order to identify and characterize biomarkers that drive the development and progression of cancer and to discover upstream genes/proteins, which could be useful to detect earlystage cancer, predict prognosis, and determine therapy efcacy or to be novel drug targets.
