Detection and quantification of post-translational modifications in non-invasive samples : Phosphoproteins as biomarkers and a market analysis of protein quantification technologies

Abstract: Post-translational modifications (PTMs) of proteins can be a sign and/or cause of disease. These modified proteins have the potential to be used as biomarkers for diagnostic purposes. However, research in the field is limited. The challenge of having an accessible way of diagnosing patients in time and at a low cost is crucial to improve public health. Blood samples or other non-invasive methods to detect diseases such as Alzheimer’s disease, Parkinson’s disease, Amyotrophic lateral sclerosis and cancers are of urgent need. This report investigates PTMs as possible biomarkers measurable in biofluids, such as blood, for diagnosis and prognosis. Biomarkers like phospho-tau and amyloid-beta are examined in the context of neurodegenerative diseases, as well as phosphorylations on neurofilaments, TAR DNA-binding protein 43 and α-synuclein. All of these are detectable in blood. Several PTMs with connection to different types of cancers are also investigated, such as F3-phosphopeptide and AFP-L3. It was found that many biomarkers for the detection of cancers can potentially be found in extracellular vesicles in blood. Methods such as ELISA, PEA, SomaScan, xMAP, SIMOA and mass spectrometry (MS) are all now available on the market to quantify these PTMs. MS has revolutionized the fields of protein detection in the past and has further evolved to being capable of protein quantification. ELISA has been prevalent for decades and laid the groundwork for improved methods such as xMAP and SIMOA that are easy to use and provide adequate sensitivity. SomaScan and PEA lead the way in dynamic range and multiplexing capacity with around 7000 and 3000 protein assays. The soon-to-be-released technology NULISA, with promising values in sensitivity and dynamic range, is also investigated here. Additionally, a written ethical analysis regarding the process and consequences of biomarker quantification through these technologies was performed. Although the investigated biomarkers are detectable in biofluids, using them as clinical diagnostic markers still poses a challenge, which is why further research in the field is needed. Through an increased knowledge of PTMs of proteins and the right use of platforms, clinical diagnostics and population screenings can be done more efficiently improving public health around the world.