Open AccessMapping of antigenic epitopes within the recombinant human preproinsulin related to insulin‐dependent diabetes mellitus
Author(s) -
Walter Michael,
Berg Heike,
Northemann Wolfgang
Publication year - 1994
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860080309
Subject(s) - recombinant dna , epitope , autoantibody , peptide , antigen , affinity chromatography , signal peptide , biochemistry , chemistry , escherichia coli , antibody , microbiology and biotechnology , biology , immunology , enzyme , gene
The human insulin domains, signal peptide, B‐chain, C‐peptide, and A‐chain, were highly expressed in Escherichia coli as recombinant proteins N‐terminally fused to glutathione‐S‐transferase and a histidinehexapeptide. The recombinant proteins were purified from insoluble cell fraction by affinity chromatography using metal chelating matrix, which was charged with Ni +2 iones. ELISA screening for autoantibodies directed to preproinsulin were performed with sera from patients with recently diagnosed insulindependent diabetes mellitus in order to localize the antigenic epitopes within the human preproinsulin. Of the patients, 14% had developed autoantibodies that recognized either the recombinant C‐peptide or the signal peptide. No reaction was observed with the A‐chain or B‐chain. © 1994 Wiley‐Liss, Inc.