Facile Preparation of Fluorescent Neoglycoproteins Using p-Nitrophenyl Anthranilate as a Heterobifunctional Linker

详细信息    查看全文

• 作者：Anthony Luyai ; Yi Lasanajak ; David F. Smith ; Richard D. Cummings ; Xuezheng Song
• 刊名：Bioconjugate Chemistry
• 出版年：2009
• 出版时间：August 19, 2009
• 年：2009
• 卷：20
• 期：8
• 页码：1618-1624
• 全文大小：395K
• 年卷期：v.20,no.8(August 19, 2009)
• ISSN：1520-4812

文摘

A facile preparation of neoglycoconjugates has been developed with a commercially available chemical, p-nitrophenyl anthranilate (PNPA), as a heterobifunctional linker. The two functional groups of PNPA, the aromatic amine and the p-nitrophenyl ester, are fully differentiated to selectively conjugate with glycans and other biomolecules containing nucleophiles. PNPA is efficiently conjugated with free reducing glycans via reductive amination. The glycan−PNPA conjugates (GPNPAs) can be easily purified and quantified by UV absorption. The active p-nitrophenyl ester in the GPNPA conjugates readily reacts with amines under mild conditions, and the resulting conjugates acquire strong fluorescence. This approach was used to prepare several fluorescent neoglycoproteins. The neoglycoproteins were covalently printed on activated glass slides and were bound by appropriate lectins recognizing the glycans.