A unique multiplex assay to profile growth factors and cytokines in CSF of MS and ALS patients

Abstract

Abstract Growth factors and cytokines play a critical role in the pathogenesis of multiple sclerosis (MS) and amyotrophic lateral sclerosis (ALS). The shift in the balance between pro-inflammatory and anti-inflammatory cytokines contributes toward autoimmunity. Epidermal growth factor (EGF) and fibroblast growth factor (FGF) are elevated in the cerebrospinal fluid (CSF) of MS and ALS patients. Identification of relevant biomarkers helps in the understanding of the disease pathogenesis. The goal is to establish a sandwich ELISA-based multiplex assay to assess growth factor and cytokine levels in CSF samples from MS and ALS patients. Capture antibodies against EGF, FGF, IFN-gamma, IL-6, IL-1beta and IL-4 imprinted in a multiplex array format into each well of a 96-well plate. Arrays subjected to multiplex sandwich ELISA of CSF samples. Test samples included CSF from Relapsing-Remitting MS (RRMS, N = 10), Secondary Progressive MS (SPMS, N = 10) and ALS patients [N=10]. Control samples included CSF from healthy subjects [N=10]. Colorimetric read-outs from multiplex ELISA are quantified using ImageJ analysis of standards and samples. Results suggest that multiplex array designed for ELISA is a unique, robust, and a cost-effective method for profiling growth factors and cytokines present in biological matrices. [*In memoriam: abstract submitted as a tribute to honor <underline>Dr. Barbara Tomik</underline> for her valuable contributions towards the collection and preparation of patient samples].