Recombinant antigens by fusion of antigenic epitopes to a GST partner

J. Molnár, I. Marczinovits, M. Kiss, S. Husz, G. Tóth, L. Dorgai, M. Kálmán

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The native human bullous pemphigoid autoantigens with molecular masses of 230 kDa and 180 kDa (BPAG1 and BPAG2, respectively) are recently used in the clinical practice by Western blot analysis for the differential diagnosis of a large group of autoimmune blistering skin diseases. Our aim was to construct ELISA and rapid diagnostic test systems with the application of 1 or 2 characteristic antigenic epitopes in the form of recombinant fusion polypeptides as low-cost and easily applicable variants of the above autoantigens. With the help of computer programmes three antigenic epitopes of the BPAG1 and BPAG2 were predicted. The chemically synthesised antigenic epitope peptides were tested by the patients sera. DNA sequences coding for the epitopes were chemically synthesised and introduced as monomer, homologous and/or heterologous dimer and multimer blocks into fusion-expression plasmids of pGEX-4T-2 (Pharmacia). The recombinant products following induction were purified from the lysates of Escherichia coli DH5α cells by affinity chromatography (Glutathione Sepharose) and used for the detection of autoantibodies of patients' sera suffering from bullous pemphigoid in an ELISA and a rapid diagnostic test. Up to 90% of 51 patients showed positive reactions with the above methods and gave far better and reliable results than were achieved by the Western blot technique with native autoantigens. We suppose that the "avidity effect" of the fusion products has greatly contributed to the high sensitivity and fidelity of the diagnostic systems developed in our laboratory.

Original languageEnglish
Pages (from-to)691-696
Number of pages6
JournalProgress in Biotechnology
Volume15
Issue numberC
DOIs
Publication statusPublished - 1998

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Epitopes
Autoantigens
Antigens
Bullous Pemphigoid
Routine Diagnostic Tests
Western Blotting
Enzyme-Linked Immunosorbent Assay
Peptides
Serum
Affinity Chromatography
Population Groups
Psychological Stress
Skin Diseases
Autoantibodies
Sepharose
Glutathione
Plasmids
Differential Diagnosis
Software
Escherichia coli

ASJC Scopus subject areas

  • Biotechnology

Cite this

Recombinant antigens by fusion of antigenic epitopes to a GST partner. / Molnár, J.; Marczinovits, I.; Kiss, M.; Husz, S.; Tóth, G.; Dorgai, L.; Kálmán, M.

In: Progress in Biotechnology, Vol. 15, No. C, 1998, p. 691-696.

Research output: Contribution to journalArticle

Molnár, J. ; Marczinovits, I. ; Kiss, M. ; Husz, S. ; Tóth, G. ; Dorgai, L. ; Kálmán, M. / Recombinant antigens by fusion of antigenic epitopes to a GST partner. In: Progress in Biotechnology. 1998 ; Vol. 15, No. C. pp. 691-696.
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