Mapping of antigenic epitopes within the recombinant human preproinsulin related to insulin-dependent diabetes mellitus
✍ Scribed by Michael Walter; Heike Berg; Dr. Wolfgang Northemann
- Publisher
- John Wiley and Sons
- Year
- 1994
- Tongue
- English
- Weight
- 591 KB
- Volume
- 8
- Category
- Article
- ISSN
- 0887-8013
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
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.