Measurement of ERK1/2 isoform distribution in peripheral blood mononuclear cells derived from whole blood from patients with different neuropsychiatric disorders

The extracellular signal regulated kinases ERK1/2 play important roles in the regulation of diverse cellular functions including proliferation, survival and apoptosis. They have been implicated in several human diseases, e. g. Alzheimerʼs disease, Parkinsonʼs disease and amyotrophic lateral sclerosis. In addition to the fully activated, diphosphorylated ERK1/2 protein, monophosphorylated forms of ERK1/2 have been observed, which may have distinct biological functions. Capillary isoelectric focusing followed by immunological detection (CIEF-Immunoassay) on a NanoPro device enables highly sensitive detection and differentiation of unphosphorylated, threonine-phosphorylated (pT), tyrosine-phosphorylated (pY) and diphosphorylated ERK1 and ERK2. We applied the technology to a pilot study addressing ERK1/2 activation in peripheral blood mononuclear cells (PBMCs) in a small cohort of patients with different neuropsychiatric disorders. PBMCs derived from whole blood of these patients were stimulated with N-methyl-D-aspartate in a time dependent manner, lysed and subjected to CIEF-Immunoassay.