Mapping of antigenic epitopes within the recombinant human preproinsulin related to insulin-dependent diabetes mellitus.
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 histidine-hexapeptide. The recombinant proteins were purified from insoluble cell fraction by affinity chromatography using metal chelating matrix, which was charged with Ni+2 ions. ELISA screening for autoantibodies directed to preproinsulin were performed with sera from patients with recently diagnosed insulin-dependent 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.
Walter
M
M
Department of Molecular Biology, ELIAS Entwicklungslabor, Freiburg, Germany.
Berg
H
H
Northemann
W
W
D001323:Autoantibodies / Q000097:blood*; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D007328:Insulin; D011384:Proinsulin / Q000276:immunology*; D011498:Protein Precursors / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
D001323:Autoantibodies; D000939:Epitopes; D007328:Insulin; D011498:Protein Precursors; D011994:Recombinant Proteins; C013737:preproinsulin; D011384:Proinsulin