Autodisplay of 60-kDa Ro/SS-A antigen and development of a surface display enzyme-linked immunosorbent assay for systemic lupus erythematosus patient sera screening

Klaudia Petermann, Stefan Vordenbäumen, Jae Chul Pyun, Achim Braukmann, Ellen Bleck, Matthias Schneider, Joachim Jose

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Enzyme-linked immunosorbent assay (ELISA) is a common tool to test human sera on an antibody reaction against a specific antigen. The 60-kDa Ro/SS-A antigen for autoantibodies can be found in sera from systemic lupus erythematosus (SLE) patients. As in the case of 60-kDa Ro/SS-A, antigens used in ELISAs are recombinantly expressed in Escherichia coli and time-consuming purification steps are needed to get the proteins. To avoid these disadvantages, 60-kDa Ro/SS-A was expressed on the surface of E. coli using autodisplay, an efficient surface display system. Cells displaying 60-kDa Ro/SS-A on the surface were applied as an antigen source instead of the purified antigen. In total, 39 patients and 30 control sera were screened on a 60-kDa Ro/SS-A antibody reaction. To eliminate antibodies against native E. coli, human sera were preabsorbed with E. coli cells prior to the assay. The new ELISA protocol (surface display ELISA [SD-ELISA]) using E. coli with autodisplayed 60-kDa Ro/SS-A showed a sensitivity of 86.67% and a specificity of 83.33% by a cutoff value of 0.28. Our results show that autodisplay provides simple, rapid, and cheap access to human antigens for an ELISA to screen human sera against specific antibody reactions.

Original languageEnglish
Pages (from-to)72-78
Number of pages7
JournalAnalytical Biochemistry
Volume407
Issue number1
DOIs
Publication statusPublished - 2010 Dec 1

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Immunosorbents
Systemic Lupus Erythematosus
Escherichia coli
Assays
Screening
Enzyme-Linked Immunosorbent Assay
Display devices
Enzymes
Antigens
Serum
Antibodies
Autoantibodies
Purification
SS-A antigen
Proteins

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Petermann, Klaudia ; Vordenbäumen, Stefan ; Pyun, Jae Chul ; Braukmann, Achim ; Bleck, Ellen ; Schneider, Matthias ; Jose, Joachim. / Autodisplay of 60-kDa Ro/SS-A antigen and development of a surface display enzyme-linked immunosorbent assay for systemic lupus erythematosus patient sera screening. In: Analytical Biochemistry. 2010 ; Vol. 407, No. 1. pp. 72-78.
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Autodisplay of 60-kDa Ro/SS-A antigen and development of a surface display enzyme-linked immunosorbent assay for systemic lupus erythematosus patient sera screening. / Petermann, Klaudia; Vordenbäumen, Stefan; Pyun, Jae Chul; Braukmann, Achim; Bleck, Ellen; Schneider, Matthias; Jose, Joachim.

In: Analytical Biochemistry, Vol. 407, No. 1, 01.12.2010, p. 72-78.

Research output: Contribution to journalArticle

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