Walter M, Berg H, Northemann W
Department of Molecular Biology, ELIAS Entwicklungslabor, Freiburg, Germany.
J Clin Lab Anal. 1994;8(3):163-6. doi: 10.1002/jcla.1860080309.
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.
人胰岛素结构域、信号肽、B链、C肽和A链在大肠杆菌中作为与谷胱甘肽-S-转移酶和组氨酸六肽N端融合的重组蛋白进行了高效表达。重组蛋白通过使用负载Ni²⁺离子的金属螯合基质的亲和色谱法从不溶性细胞部分中纯化出来。为了确定人胰岛素原中的抗原表位，使用近期诊断为胰岛素依赖型糖尿病患者的血清进行了针对胰岛素原的自身抗体的ELISA筛选。在这些患者中，14%产生了识别重组C肽或信号肽的自身抗体。未观察到与A链或B链的反应。
