Affinity purification and comparative biosensor analysis of citrulline-peptide-specific antibodies in rheumatoid arthritis

Eszter Szarka, Petra Aradi, Krisztina Huber, Judit Pozsgay, Lili Végh, Anna Magyar, Gergő Gyulai, György Nagy, Bernadette Rojkovich, E. Kiss, F. Hudecz, G. Sármay

Research output: Contribution to journalArticle

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 varied between 10−6 and 10−8 M and inversely correlated 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 detected antibodies in RA sera with 66% sensitivity and 98% specificity in ELISA and was recognized by 90% of RA sera, while none of the healthy samples in SPR. When coupled to nanoparticles, the multi-epitope peptide specifically targeted and depleted ACPA-producing B cells ex vivo. Conclusions: The unique multi-epitope peptide designed based on ACPA cross-reactivity might be suitable to develop better diagnostics and novel therapies for RA. View Full-Text.

Original languageEnglish
Article number326
JournalInternational Journal of Molecular Sciences
Volume19
Issue number1
DOIs
Publication statusPublished - Jan 22 2018

Fingerprint

arthritis
Citrulline
Biosensing Techniques
antibodies
bioinstrumentation
Biosensors
purification
Antibodies
Peptides
peptides
Purification
affinity
Rheumatoid Arthritis
Epitopes
proteins
Proteins
serums
Antibody-Producing Cells
Surface Plasmon Resonance
Surface plasmon resonance

Keywords

  • Affinity
  • Autoantibody
  • Citrulline-peptides
  • Cross-reaction
  • Rheumatoid arthritis
  • Targeting

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

Cite this

Affinity purification and comparative biosensor analysis of citrulline-peptide-specific antibodies in rheumatoid arthritis. / Szarka, Eszter; Aradi, Petra; Huber, Krisztina; Pozsgay, Judit; Végh, Lili; Magyar, Anna; Gyulai, Gergő; Nagy, György; Rojkovich, Bernadette; Kiss, E.; Hudecz, F.; Sármay, G.

In: International Journal of Molecular Sciences, Vol. 19, No. 1, 326, 22.01.2018.

Research output: Contribution to journalArticle

Szarka, Eszter ; Aradi, Petra ; Huber, Krisztina ; Pozsgay, Judit ; Végh, Lili ; Magyar, Anna ; Gyulai, Gergő ; Nagy, György ; Rojkovich, Bernadette ; Kiss, E. ; Hudecz, F. ; Sármay, G. / Affinity purification and comparative biosensor analysis of citrulline-peptide-specific antibodies in rheumatoid arthritis. In: International Journal of Molecular Sciences. 2018 ; Vol. 19, No. 1.
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AU - Szarka, Eszter

AU - Aradi, Petra

AU - Huber, Krisztina

AU - Pozsgay, Judit

AU - Végh, Lili

AU - Magyar, Anna

AU - Gyulai, Gergő

AU - Nagy, György

AU - Rojkovich, Bernadette

AU - Kiss, E.

AU - Hudecz, F.

AU - Sármay, G.

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N2 - 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 varied between 10−6 and 10−8 M and inversely correlated 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 detected antibodies in RA sera with 66% sensitivity and 98% specificity in ELISA and was recognized by 90% of RA sera, while none of the healthy samples in SPR. When coupled to nanoparticles, the multi-epitope peptide specifically targeted and depleted ACPA-producing B cells ex vivo. Conclusions: The unique multi-epitope peptide designed based on ACPA cross-reactivity might be suitable to develop better diagnostics and novel therapies for RA. View Full-Text.

AB - 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 varied between 10−6 and 10−8 M and inversely correlated 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 detected antibodies in RA sera with 66% sensitivity and 98% specificity in ELISA and was recognized by 90% of RA sera, while none of the healthy samples in SPR. When coupled to nanoparticles, the multi-epitope peptide specifically targeted and depleted ACPA-producing B cells ex vivo. Conclusions: The unique multi-epitope peptide designed based on ACPA cross-reactivity might be suitable to develop better diagnostics and novel therapies for RA. View Full-Text.

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