On-Array Citrullination of Protein Microarrays.

Area of Science:

• Biochemistry
• Immunology
• Molecular Biology

Background:

• Protein microarrays are crucial for studying autoimmune diseases.
• Autoantibody responses often target modified proteins (neo-autoantigens), not native ones.
• Citrullination, a post-translational modification, is central to rheumatoid arthritis (RA) pathogenesis.

Purpose of the Study:

• To describe a protocol for on-array citrullination of protein microarrays.
• To replicate in vivo citrullination processes associated with RA.
• To enable high-throughput identification of citrullinated autoantigens.

Main Methods:

• Incubation of protein microarrays with a peptidylarginine deiminase (PAD) containing buffer.
• On-array citrullination to generate citrullinated proteins.
• Utilizing protein microarrays for autoantigen discovery.

Main Results:

• The protocol successfully performs on-array citrullination, mimicking in vivo conditions.
• This method facilitates the identification of citrullinated autoantigens.
• Provides a platform for discovering novel RA autoantigens.

Conclusions:

• On-array citrullination is an effective strategy for generating and identifying neo-autoantigens.
• This approach enhances the study of autoimmune disease pathogenesis, particularly RA.
• Facilitates the discovery of novel autoantigens and their roles in disease.