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首页> 外文期刊>Journal of peptide science: An official publication of the European Peptide Society >Easy parallel screening of reagent stability, quality control, and metrology in solid phase peptide synthesis (SPPS) and peptide couplings for microarrays
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Easy parallel screening of reagent stability, quality control, and metrology in solid phase peptide synthesis (SPPS) and peptide couplings for microarrays

机译：轻松并行筛选固相肽合成（SPPS）和用于微阵列的肽偶联中的试剂稳定性，质量控制和计量学

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Evaluating the stability of coupling reagents, quality control (QC), and surface functionalization metrology are all critical to the production of high quality peptide microarrays. We describe a broadly applicable screening technique for evaluating the fidelity of solid phase peptide synthesis (SPPS), the stability of activation/coupling reagents, and a microarray surface metrology tool. This technique was used to assess the stability of the activation reagent 1-{[1-(Cyano-2-ethoxy-2-oxo-ethylidenaminooxy)dimethylamino-morpholinomethylene]} methaneaminiumHexafluorophosphate (COMU) (Sigma-Aldrich, St. Louis, MO, USA) by SPPS of Leu-Enkephalin (YGGFL) or the coupling of commercially synthesized YGGFL peptides to (3-aminopropyl) triethyoxysilane-modified glass surfaces. Coupling efficiency was quantitated by fluorescence signaling based on immunoreactivity of the YGGFL motif. It was concluded that COMU solutions should be prepared fresh and used within 5 h when stored at similar to 23 degrees C and not beyond 24 h if stored refrigerated, both in closed containers. Caveats to gauging COMU stability by absorption spectroscopy are discussed. Commercial YGGFL peptides needed independent QC, due to immunoreactivity variations for the same sequence synthesized by different vendors. This technique is useful in evaluating the stability of other activation/coupling reagents besides COMU and as a metrology tool for SPPS and peptide microarrays. Copyright (C) 2015 European Peptide Society and John Wiley & Sons, Ltd.
机译：评估偶联试剂的稳定性，质量控制（QC）和表面功能化计量对于生产高质量肽微阵列都是至关重要的。我们描述了一种广泛适用的筛选技术，用于评估固相肽合成（SPPS）的保真度，激活/偶联试剂的稳定性以及微阵列表面计量工具。该技术用于评估活化试剂1-{[1-（氰基-2-乙氧基-2-氧代-亚乙基氨基氧基）二甲基氨基-吗啉代亚甲基]}甲铵六氟磷酸酯（COMU）的稳定性（Sigma-Aldrich，St.Louis，MO （美国）通过Leu-Enkephalin（YGGFL）的SPPS或将商业合成的YGGFL肽偶联到（3-氨丙基）三乙氧基硅烷改性的玻璃表面上。偶联效率通过基于YGGFL基序的免疫反应性的荧光信号定量。得出的结论是，COMU解决方案应在接近23摄氏度的条件下新鲜制备并在5小时内使用，并且如果冷藏保存，则应在密闭容器中保存24小时以内。讨论了通过吸收光谱法测量COMU稳定性的注意事项。由于不同供应商合成的同一序列的免疫反应性差异，商业YGGFL肽需要独立的QC。该技术可用于评估COMU以外的其他激活/偶联试剂的稳定性，并可用作SPPS和肽微阵列的计量工具。版权所有（C）2015欧洲多肽协会和John Wiley＆Sons，Ltd.

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