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Optimization of Solid-Phase Synthesis of the 1-40 Beta-Amyloid and Preparation of Antibodies Revealing It under Immunoblotting Conditions
Russian Journal of Bioorganic Chemistry ( IF 1 ) Pub Date : 2020-03-01 , DOI: 10.1134/s1068162020020181 T. D. Volkova , D. O. Koroev , A. V. Kamynina , M. P. Filatova , A. V. Avetisyan , O. M. Volpina
Russian Journal of Bioorganic Chemistry ( IF 1 ) Pub Date : 2020-03-01 , DOI: 10.1134/s1068162020020181 T. D. Volkova , D. O. Koroev , A. V. Kamynina , M. P. Filatova , A. V. Avetisyan , O. M. Volpina
Abstract The solid-phase synthesis and purification of the 1-40 sequence of the human beta-amyloid were optimized, resulting in a preparation of a product with a high yield and homogeneity more than 95%. The synthetic peptide is capable of forming oligomers. This fact was confirmed by electrophoresis in the polyacrylamide gel with a subsequent immunoblotting and fluorescence spectrophotometry using the thioflavin T dye. An available method for a production of the highly specific anti-beta-amyloid antibodies with a high titer was developed. These antibodies recognized both monomeric and oligomeric forms of the 1-40 peptide of beta-amyloid under the immunoblotting conditions.
中文翻译:
1-40 β-淀粉样蛋白的固相合成优化和在免疫印迹条件下揭示它的抗体的制备
摘要 对人β-淀粉样蛋白1-40序列的固相合成和纯化进行了优化,制备的产物收率高,均一性超过95%。合成肽能够形成寡聚体。这一事实通过聚丙烯酰胺凝胶电泳以及随后使用硫代黄素 T 染料的免疫印迹和荧光分光光度法得到证实。开发了一种生产具有高滴度的高特异性抗β-淀粉样蛋白抗体的可用方法。这些抗体在免疫印迹条件下识别 β-淀粉样蛋白的 1-40 肽的单体和寡聚形式。
更新日期:2020-03-01
中文翻译:
1-40 β-淀粉样蛋白的固相合成优化和在免疫印迹条件下揭示它的抗体的制备
摘要 对人β-淀粉样蛋白1-40序列的固相合成和纯化进行了优化,制备的产物收率高,均一性超过95%。合成肽能够形成寡聚体。这一事实通过聚丙烯酰胺凝胶电泳以及随后使用硫代黄素 T 染料的免疫印迹和荧光分光光度法得到证实。开发了一种生产具有高滴度的高特异性抗β-淀粉样蛋白抗体的可用方法。这些抗体在免疫印迹条件下识别 β-淀粉样蛋白的 1-40 肽的单体和寡聚形式。