量化α-苯基-D-甘露糖胺对癌细胞表面唾液酸进行代谢工程改造的效率。
Quantifying the Efficiency of -Phenyl-D-mannosamine to Metabolically Engineer Sialic Acid on Cancer Cell Surface.
作者信息
Zhou Zhifang, Liao Guochao, Stepanovs Sergejs, Guo Zhongwu
机构信息
Department of Chemistry, Wayne State University, Detroit, Michigan 48202, USA.
出版信息
J Carbohydr Chem. 2014 Nov 1;33(7-8):395-407. doi: 10.1080/07328303.2014.933483.
A convenient method was developed for the quantification of sialic acids expressed by cells and used to analyze the efficiency of -phenylacetyl-D-mannosamine (ManNPhAc) to metabolically glycoengineer SKMEL-28 cancer cell. For this purpose, ManNPhAc-cultured cells were treated with 2M acetic acid to release sialic acids, and the products were treated with 1,2-diamino-4,5-methylenedioxybenzene to form the corresponding derivatives that had strong UV absorptions. The reaction mixture was then applied to HPLC-UV analysis to determine the amounts and the ratios of natural sialic acid and its unnatural analog. It was confirmed that after incubation with ManNPhAc SKMEL-28 cell was effectively glycoengineered to express a significant amount of unnatural sialic acid.
开发了一种用于定量细胞表达的唾液酸的简便方法，并用于分析苯乙酰-D-甘露糖胺（ManNPhAc）对SKMEL-28癌细胞进行代谢糖工程改造的效率。为此，用2M乙酸处理经ManNPhAc培养的细胞以释放唾液酸，产物用1,2-二氨基-4,5-亚甲二氧基苯处理以形成具有强紫外吸收的相应衍生物。然后将反应混合物用于HPLC-UV分析，以确定天然唾液酸及其非天然类似物的量和比例。证实与ManNPhAc孵育后，SKMEL-28细胞被有效地进行了糖工程改造，以表达大量的非天然唾液酸。
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