Diagnostics of autoimmune neurodegeneration using fluorescent probing

Scientific Reports
Yakov LomakinA A Belogurov

Abstract

The discovery of antibody-mediated catalysis was a breakthrough that showed antibody function is not limited to specific binding interactions, and that immunoglobulins (Igs) may also chemically transform their target antigens. Recently, so-called "natural catalytic antibodies" have been intimately linked with several pathologies, where they either protect the organism or contribute to the development of autoimmune abnormalities. Previously, we showed that myelin-reactive autoantibodies from patients with multiple sclerosis (MS) and mice with experimental autoimmune encephalomyelitis (EAE) exhibit the ability to recognize and hydrolyse distinct epitopes within myelin basic protein (MBP). Further, the antibody-mediated cleavage of encephalitogenic MBP peptide 81-103, flanked by two fluorescent proteins, can serve as a novel biomarker for MS. Here, we report the next generation of this biomarker, based on the antibody-mediated degradation of a novel chemically synthesized FRET substrate, comprising the fluorophore Cy5 and the quencher QXL680, interconnected by the MBP peptide 81-99: Cy5-MBP81-99-QXL680. This substrate is degraded upon incubation with either purified antibodies from MS patients but not healthy donors or purified an...Continue Reading

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Methods Mentioned

BETA
density gradient centrifugation
FRET
biosensor
gel-filtration
electrophoresis
size-exclusion chromatography
ELISA
reverse-phase chromatography

Software Mentioned

Sigma
Plot
SPSS

Related Concepts

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