TY - JOUR
T1 - A contra capture protein array platform for studying post-translationally modified (PTM) auto-antigenomes
AU - Karthikeyan, Kailash
AU - Barker, Kristi
AU - Tang, Yanyang
AU - Kahn, Peter
AU - Wiktor, Peter
AU - Brunner, Al
AU - Knabben, Vinicius
AU - Takulapalli, Bharath
AU - Buckner, Jane
AU - Nepom, Gerald
AU - LaBaer, Joshua
AU - Qiu, Ji
N1 - Funding Information:
This work was funded by the National Institutes of Health (NIH) grant R21 (ZDS0078), partially by National Institutes of Health (NIH) grant R42GM106704 awarded to Engineering Arts LLC. and Virginia G. Piper Center for Personalized Diagnostics at the Biodesign Institute (Arizona State University).
Publisher Copyright:
© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2016/7
Y1 - 2016/7
N2 - Aberrant modifications of proteins occur during disease development and elicit disease-specific antibody responses. We have developed a protein array platform that enables the modification of many proteins in parallel and assesses their immunogenicity without the need to express, purify, and modify proteins individually. We used anticitrullinated protein antibodies (ACPAs) in rheumatoid arthritis (RA) as a model modification and profiled antibody responses to 190 citrullinated proteins in 20 RA patients. We observed unique antibody reactivity patterns in both clinical anticyclic citrullinated peptide assay positive (CCP) and CCP-RA patients. At individual antigen levels, we detected antibodies against known citrullinated autoantigens and discovered and validated five novel antibodies against specific citrullinated antigens (osteopontin (SPP1), flap endonuclease (FEN1), insulin like growth factor binding protein 6 (IGFBP6), insulin like growth factor I (IGF1) and stanniocalcin-2 (STC2)) in RA patients. We also demonstrated the utility of our innovative array platform in the identification of immune-dominant epitope(s) for citrullinated antigens. We believe our platform will promote the study of post-translationally modified antigens at a breadth that has not been achieved before, by both identifying novel autoantigens and investigating their roles in disease development. The developed platforms can potentially be used to study many autoimmune disease-relevant modifications and their immunogenicity.
AB - Aberrant modifications of proteins occur during disease development and elicit disease-specific antibody responses. We have developed a protein array platform that enables the modification of many proteins in parallel and assesses their immunogenicity without the need to express, purify, and modify proteins individually. We used anticitrullinated protein antibodies (ACPAs) in rheumatoid arthritis (RA) as a model modification and profiled antibody responses to 190 citrullinated proteins in 20 RA patients. We observed unique antibody reactivity patterns in both clinical anticyclic citrullinated peptide assay positive (CCP) and CCP-RA patients. At individual antigen levels, we detected antibodies against known citrullinated autoantigens and discovered and validated five novel antibodies against specific citrullinated antigens (osteopontin (SPP1), flap endonuclease (FEN1), insulin like growth factor binding protein 6 (IGFBP6), insulin like growth factor I (IGF1) and stanniocalcin-2 (STC2)) in RA patients. We also demonstrated the utility of our innovative array platform in the identification of immune-dominant epitope(s) for citrullinated antigens. We believe our platform will promote the study of post-translationally modified antigens at a breadth that has not been achieved before, by both identifying novel autoantigens and investigating their roles in disease development. The developed platforms can potentially be used to study many autoimmune disease-relevant modifications and their immunogenicity.
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U2 - 10.1074/mcp.M115.057661
DO - 10.1074/mcp.M115.057661
M3 - Article
C2 - 27141097
AN - SCOPUS:84977264250
SN - 1535-9476
VL - 15
SP - 2324
EP - 2337
JO - Molecular and Cellular Proteomics
JF - Molecular and Cellular Proteomics
IS - 7
ER -