Quantifying the Efficiency of N-Phenyl-D-mannosamine to Metabolically Engineer Sialic Acid on Cancer Cell Surface

A convenient method was developed for the quantification of sialic acids expressed by cells and used to analyze the efficiency of N-phenylacetyl-D-mannosamine (ManNPhAc) to metabolically glycoengineer SKMEL-28 cancer cells. 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, the SKMEL-28 cell was effectively glycoengineered to express a significant amount of unnatural sialic acid.