Mimotopes for lupus-derived anti-DNA and nucleosome-specific autoantibodies selected from random peptide phage display libraries: Facts and follies

Jürgen W. Dieker, Yong Jiang Sun, Cor W. Jacobs, Chaim Putterman, Marc Monestier, Sylviane Muller, Johan Van Der Vlag, Jo H. Berden

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Autoantibodies against chromatin are the most characteristic serological feature in SLE patients. Anti-dsDNA and nucleosome-specific antibodies are associated with glomerulonephritis, the most serious manifestation of SLE. Identification of peptides mimicking conformational epitopes (so-called mimotopes) on the nucleosome recognized by these antibodies is of considerable interest. Using an approach similar to that used previously to characterize mimotopes for anti-DNA autoantibodies, we have selected and identified a mimotope for a nucleosome-specific autoantibody (#32) by screening a random peptide phage display library. However, the reactivity of monoclonal antibody (mAb) #32 with the selected mimotope (MIMO#0) in ELISA was dependent on the blocking reagents used. Using nonfat dry milk (5%), mAb #32 clearly bound to MIMO#0, but using fetal bovine calf serum (FCS) (5%), there was no binding. Furthermore, again dependent on the blocking reagent used in ELISA, the selected mimotope MIMO#0 was not only recognized by the selecting antibody mAb #32, but also by a large number of other monoclonal anti-DNA, anti-histone and nucleosome-specific autoantibodies (NSA). We could demonstrate that the selected mimotope was able to bind directly to nucleosomal material (DNA/histone complexes) and labeled DNA. This finding was extended to other previously identified mimotopes for anti-DNA autoantibodies. We conclude that nucleosomal material (DNA/histone complexes), derived from reagents used during the mimotope selection procedure, resulted in the selection of DNA-binding peptides from the phage display library, rather than mimotopes. In addition, we could demonstrate that blocking reagents greatly influence the reactivity of anti-DNA, anti-histone and nucleosome-specific autoantibodies in ELISA. Development of blocking reagents devoid of nucleosomal material (DNA/histone complexes) is urgently needed for assay systems in which anti-nuclear autoantibodies are tested.

Original languageEnglish (US)
Pages (from-to)83-93
Number of pages11
JournalJournal of Immunological Methods
Volume296
Issue number1-2
DOIs
StatePublished - Jan 2005

Fingerprint

Peptide Library
Nucleosomes
Autoantibodies
DNA
Histones
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Antibodies
Glomerulonephritis
Chromatin
Epitopes
Milk
Peptides

Keywords

  • Autoantibodies
  • Mimotopes
  • Random peptide phage display
  • Systemic lupus erythematosus

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Mimotopes for lupus-derived anti-DNA and nucleosome-specific autoantibodies selected from random peptide phage display libraries : Facts and follies. / Dieker, Jürgen W.; Sun, Yong Jiang; Jacobs, Cor W.; Putterman, Chaim; Monestier, Marc; Muller, Sylviane; Van Der Vlag, Johan; Berden, Jo H.

In: Journal of Immunological Methods, Vol. 296, No. 1-2, 01.2005, p. 83-93.

Research output: Contribution to journalArticle

Dieker, Jürgen W. ; Sun, Yong Jiang ; Jacobs, Cor W. ; Putterman, Chaim ; Monestier, Marc ; Muller, Sylviane ; Van Der Vlag, Johan ; Berden, Jo H. / Mimotopes for lupus-derived anti-DNA and nucleosome-specific autoantibodies selected from random peptide phage display libraries : Facts and follies. In: Journal of Immunological Methods. 2005 ; Vol. 296, No. 1-2. pp. 83-93.
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abstract = "Autoantibodies against chromatin are the most characteristic serological feature in SLE patients. Anti-dsDNA and nucleosome-specific antibodies are associated with glomerulonephritis, the most serious manifestation of SLE. Identification of peptides mimicking conformational epitopes (so-called mimotopes) on the nucleosome recognized by these antibodies is of considerable interest. Using an approach similar to that used previously to characterize mimotopes for anti-DNA autoantibodies, we have selected and identified a mimotope for a nucleosome-specific autoantibody (#32) by screening a random peptide phage display library. However, the reactivity of monoclonal antibody (mAb) #32 with the selected mimotope (MIMO#0) in ELISA was dependent on the blocking reagents used. Using nonfat dry milk (5{\%}), mAb #32 clearly bound to MIMO#0, but using fetal bovine calf serum (FCS) (5{\%}), there was no binding. Furthermore, again dependent on the blocking reagent used in ELISA, the selected mimotope MIMO#0 was not only recognized by the selecting antibody mAb #32, but also by a large number of other monoclonal anti-DNA, anti-histone and nucleosome-specific autoantibodies (NSA). We could demonstrate that the selected mimotope was able to bind directly to nucleosomal material (DNA/histone complexes) and labeled DNA. This finding was extended to other previously identified mimotopes for anti-DNA autoantibodies. We conclude that nucleosomal material (DNA/histone complexes), derived from reagents used during the mimotope selection procedure, resulted in the selection of DNA-binding peptides from the phage display library, rather than mimotopes. In addition, we could demonstrate that blocking reagents greatly influence the reactivity of anti-DNA, anti-histone and nucleosome-specific autoantibodies in ELISA. Development of blocking reagents devoid of nucleosomal material (DNA/histone complexes) is urgently needed for assay systems in which anti-nuclear autoantibodies are tested.",
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