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Generation of Two-color Antigen Microarrays for the Simultaneous Detection of IgG and IgM Autoantibodies

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Published on: September 15, 2016

Caspr2 autoantibodies target multiple epitopes.

Abby L Olsen¹, Yongjie Lai¹, Josep Dalmau¹

¹Department of Neurology (A.L.O., Y.L., J.D., S.S.S., E.L.), The University of Pennsylvania, Philadelphia; and ICREA-IDIBAPS (J.D.), Hospital Unit, University of Barcelona, Spain.

Neurology(R) Neuroimmunology & Neuroinflammation

|July 18, 2015

Summary

This summary is machine-generated.

Autoantibodies targeting contactin-associated protein-like 2 (Caspr2) recognize multiple extracellular epitopes, including within the discoidin-like domain. Protein structure and glycosylation are not essential for this antibody reactivity.

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Area of Science:

Neuroimmunology
Molecular Neuroscience
Autoimmunity

Background:

Autoantibodies against contactin-associated protein-like 2 (Caspr2) are implicated in neurological disorders.
Understanding the specific target epitopes of these autoantibodies is crucial for elucidating disease mechanisms.

Purpose of the Study:

To investigate the target epitopes of autoantibodies against the axonal protein Caspr2.
To determine the role of protein structure and glycosylation in Caspr2 autoantibody recognition.

Main Methods:

Engineered Caspr2 constructs with deletions in extracellular subdomains.
Immunofluorescence staining and Western blotting using patient sera and a commercial antibody.
Assessed the impact of glycosylation inhibition on antibody binding.

Main Results:

Patient autoantibodies bound to the extracellular domain of Caspr2.
Reactivity was observed even without native protein structure or glycosylation.
Deletion of N-terminal subdomains, particularly the discoidin-like domain, significantly reduced or abolished antibody binding.

Conclusions:

Caspr2 autoantibodies target multiple epitopes within the extracellular domain.
The discoidin-like domain contains a significant epitope for Caspr2 autoantibodies.
Glycosylation and native protein conformation are not strictly required for Caspr2 autoantibody recognition.