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Immunoprecipitation

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...

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Exploring autoantigens in autoimmune limbic encephalitis using phage immunoprecipitation sequencing.

Haidara Kherbek^1,2, Naveen K Paramasivan^1,2, Surendra Dasari^1,2

¹Department of Neurology, Mayo Clinic, 200 1st Street, SW, Rochester, MN, USA.

Journal of Neurology

|March 26, 2025

Summary

High-throughput phage immunoprecipitation sequencing (PhIP-Seq) identified neuronal PAS domain protein 4 (NPAS4) as a potential autoantigen in antibody-negative autoimmune limbic encephalitis (ALE). Additional cases of adenylate kinase 5 (AK5)-IgG positive ALE were also identified, highlighting PhIP-Seq

Keywords:

Adenylate kinase 5 (AK5)Autoimmune encephalitis Cognitive dysfunction Limbic encephalitis Neuronal PAS domain protein 4 (NPAS4)Phage immunoprecipitation sequencing (PhIP-seq)

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Antibody Profiling by Luciferase Immunoprecipitation Systems LIPS

Antibody Profiling by Luciferase Immunoprecipitation Systems LIPS

Published on: October 7, 2009

Area of Science:

Neuroimmunology
Proteomics
Autoimmune Neurology

Background:

Autoimmune limbic encephalitis (ALE) can be challenging to diagnose when specific autoantibodies are absent.
Identifying novel autoantigens is crucial for understanding the pathogenesis of antibody-negative ALE.
High-throughput methods are needed to screen for potential autoantigens in complex neurological disorders.

Purpose of the Study:

To employ high-throughput whole-human proteome phage immunoprecipitation sequencing (PhIP-Seq) to discover potential autoantigens in antibody-negative ALE.
To validate identified autoantigens using serological and cellular assays.
To characterize the clinical and radiological features associated with novel autoantigen discoveries.

Main Methods:

PhIP-Seq was performed on cerebrospinal fluid (CSF) samples from patients diagnosed with antibody-negative ALE.
Putative autoantigens were validated using recombinant protein-based assays, cell-based assays (CBA), and enzyme-linked immunosorbent assays (ELISA).
A large cohort of disease and healthy controls were tested for specificity using ELISA.

Main Results:

PhIP-Seq identified neuronal PAS domain protein 4 (NPAS4) as a putative autoantigen in two ALE patients with distinct clinical and MRI findings.
NPAS4-IgG autoantibodies were confirmed by CBA and ELISA, and specificity was demonstrated against a large control cohort.
Three additional ALE patients with adenylate kinase 5 (AK5)-IgG positivity were identified, presenting with characteristic medial temporal lobe involvement and memory loss.

Conclusions:

High-throughput PhIP-Seq is a valuable tool for identifying novel autoantigens in antibody-negative ALE.
NPAS4 is identified as a potential autoantigen in a subset of ALE patients.
The findings expand the spectrum of known autoantigens in ALE and underscore the utility of PhIP-Seq in autoimmune research.