Sequencing of Complex Mixtures of Heparan Sulfate Tetrasaccharides by Chemical Derivatization and LC-MS/MS

Through a collaboration between synthetic chemists investigating chemical synthesis of defined heparan sulfate oligosaccharides (Chengli Zong, Andre Venot, and Geert-Jan Boons), analytical chemists developing a derivatization, separation, and MS/MS methods for separating and sequencing complex mixtures of heparan sulfate oligosaccharides (Rongrong Huang, Dandan Zhou, and Joshua Sharp), and a bioinformaticist developing software for high-throughput analysis of heparan sulfate sequencing results (Yulun Chiu), we have developed and demonstrated a semi-automated method capable of separating and sequencing complex mixtures of heparin and heparan sulfate, including structures that are very closely related to one another. Only through the availability of both well-defined synthetic standards and advanced analytical technologies could we develop a tool that is demonstrated to be robust and reliable. Computational tools have been developed to perform automated data analysis for high-throughput heparin and heparan sulfate sequencing. A version of this software is scheduled for public release in 2014, with periodic updates made publicly available as we develop them.