Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Affinity Purification and Comparative Biosensor Analysis of Citrulline-Peptide Specific Antibodies in Rheumatoid Arthritis

Eszter Szarka
ORCID logo ,
Petra Aradi
,
Krisztina Huber
,
Judit Pozsgay
,
Lili Végh
,
Anna Magyar
,
Gergő Gyulai
,
György Nagy
,
Bernadette Rojkovich
,
Eva Kiss
,
Ferenc Hudecz
,
Gabriella Sármay
* ORCID logo
Version 1 : Received: 30 December 2017 / Approved: 2 January 2018 / Online: 2 January 2018 (11:50:04 CET)

A peer-reviewed article of this Preprint also exists.

Szarka, E.; Aradi, P.; Huber, K.; Pozsgay, J.; Végh, L.; Magyar, A.; Gyulai, G.; Nagy, G.; Rojkovich, B.; Kiss, É.; Hudecz, F.; Sármay, G. Affinity Purification and Comparative Biosensor Analysis of Citrulline-Peptide-Specific Antibodies in Rheumatoid Arthritis. Int. J. Mol. Sci. 2018, 19, 326. Szarka, E.; Aradi, P.; Huber, K.; Pozsgay, J.; Végh, L.; Magyar, A.; Gyulai, G.; Nagy, G.; Rojkovich, B.; Kiss, É.; Hudecz, F.; Sármay, G. Affinity Purification and Comparative Biosensor Analysis of Citrulline-Peptide-Specific Antibodies in Rheumatoid Arthritis. Int. J. Mol. Sci. 2018, 19, 326.

Abstract

Background: In rheumatoid arthritis (RA), anti-citrullinated protein/peptide antibodies (ACPAs) are responsible for disease onset and progression, however, our knowledge is limited on ligand binding affinities of autoantibodies with different citrulline-peptide specificity. Methods: Citrulline-peptide specific ACPA IgGs were affinity purified and tested by ELISA. Binding affinities of ACPA IgGs and serum antibodies were compared by surface plasmon resonance (SPR) analysis. Bifunctional nanoparticles harboring a multi-epitope citrulline-peptide and a complement activating peptide were used to induce selective depletion of ACPA producing B cells. Results: KD values of affinity purified ACPA IgGs varies between 10-6-10-8 M and inversely correlate with disease activity. Based on their cross-reaction with citrulline-peptides we designed a novel multi-epitope peptide, containing Cit-Gly and Ala-Cit motifs in two-two copies, separated with a short, neutral spacer. This peptide detects antibodies in RA sera with 66 % sensitivity and 98 % specificity in ELISA and is recognized by 90% of RA sera, while none of the healthy samples in SPR. When coupled to nanoparticles, the multi-epitope peptide specifically targets and depletes ACPA producing B cells ex vivo. Conclusions: The unique multi-epitope peptide designed on the basis of ACPA cross-reactivity might be suitable to develop better diagnostics and novel therapies for RA.

Keywords

rheumatoid arthritis; citrulline-peptides; autoantibody; affinity; cross-reaction; targeting

Subject

Biology and Life Sciences, Immunology and Microbiology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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