Application of Proteomics Technology to the Field of Neurotrauma

Abstract

Near-completion of the Human Genome Project has stimulated scientists to begin looking for the next step in unraveling normal and abnormal functions within biological systems. Consequently, there is new focus on the role of proteins in these processes. Proteomics is a burgeoning field that may provide a valuable approach to evaluate the post-traumatic central nervous system (CNS). Although we cannot provide a comprehensive assessment of all methods for protein analysis, this report summarizes some of the newer proteomic technologies that have propelled this field into the limelight and that are available to most researchers in neurotrauma. Three technical approaches (two-dimensional gel electrophoresis, direct analysis by mass spectrometry, including two-dimensional chromatography coupled to mass spectrometry and isotope coded affinity tags, and antibody technologies) are reviewed, and their advantages and disadvantages presented. A discussion of proteomic technology in the context of brain and spinal cord trauma follows, addressing current and future challenges. Proteomics will likely be very useful for developing diagnostic predictors after CNS injury and for mapping changes in proteins after injury in order to identify new therapeutic targets. Neurotrauma results in complex alterations to the biological systems within the nervous system, and these changes evolve over time. Exploration of the "new nervous system" that follows injury will require methods that can both fully assess and simplify this complexity.

Post-Genomics Nanotechnology Is Gaining Momentum: Nanoproteomics and Applications in Life Sciences

“…For example, on the level of brain proteome, it is estimated that there exist around 20,000 brain proteins that are differentially expressed in the various regions of the brain (Wang et al, 2005a). Furthermore, it is difficult to associate mRNA expression to the protein expression levels (Denslow et al, 2003;Freeman et al, 2005;Morrison et al, 2002;Wang et al, 2004). This is due to several factors, including ''alternative splicing,'' which is highly common in brain tissue, generating several copies of highly related splices from a single gene (Hunnerkopf et al, 2007;Missler and Sudhof, 1998;Morrison et al, 2002;Wu and Maniatis, 1999).…”

Exploring the Role of Biomarkers for the Diagnosis and Management of Traumatic Brain Injury Patients

“…Some of the disadvantages include sample to sample variation, the inability to detect certain classes or sizes of proteins, and the need for many samples and controls. 40 There are also non-gel-based mass spectrometry methods for identifying proteins that use high-resolution chromatography to separate complex mixtures of proteins prior to mass spectrometry. Typically the technique uses capillary chromatography for sensitivity and high-resolution mass spectrometry for identification of proteins.…”

“…However, the technique requires significant expertise and the cost of the materials and equipment to run this technique is much higher. 40 Newer proteomic techniques are employing antibody-based methods such as high throughput immunoblotting and antibody panels and/or arrays (ELISA's). Antibodies are significantly more specific and selective than traditional techniques and allow the detection of proteins amid complex high-protein content biofluids such as serum or plasma.…”

Novel Strategies for Discovery, Validation and FDA Approval of Biomarkers for Acute and Chronic Brain Injury

“…Neuroproteomics and neurogenomics, the application of proteomics and genomics in the field of neuronal injury, have been identified as a potential means for biomarker discovery, with the ability to identify proteome dynamics in response to brain injury (Guingab-Cagmat 2009, Haskins et al 2005, Davidsson & Sjogren 2005, Shin et al 2002, Celis et al 2004). In the area of brain injury, several studies have demonstrated the role of proteomics (Denslow et al 2003, Katano et al 2006) and genomics (Redell et al 2009, Ding et al 2006 in providing significant insight into understanding changes, modifications and functions in certain proteins post TBI. In addition, genomics and proteomics are powerful, complementary tools that play an important role in the area of biomarker identification.…”