Chris Ashwood Chris Ashwood Ph.D., is the glycomics core director at the Beth Israel Deaconess Medical Center (BIDMC, Boston, Massachusetts). He received his Ph.D. in analytical glycobiology at Macquarie University, Australia supervised by Prof. Nicolle Packer. He joined the Glycomics Core after his postdoc work at the Medical College of Wisconsin and University of Nebraska Medical Center. The core provides services, instrumentation, and expertise in glycomics to collaborators and clients across the world.

High-throughput Glycan Composition Profiling Enabled by MALDISkyLink and the Skyline Ecosystem

Protein glycosylation is the most frequent and varied type of co- and post-translational modification seen in eukaryotic cells. Identification of the glycan compositions that modify individual proteins (e.g. therapeutic proteins) or protein mixtures (e.g. cell lysates and plasma) can open the gateway to the glycome for investigators from a range of backgrounds. Released glycans from 384-sample plates can be rapidly profiled by MALDI-TOF-MS, generating a single representative spectrum every 10 seconds. Despite the acquisition speed, data analysis throughput is lacking. Using our newly developed software, MALDISkyLink, individual spectra are converted into a format compatible with the Skyline ecosystem, which is typically limited to LC- or CE- MS data. This enables the use of Skyline Batch for MALDI-based glycan analysis by extracting precursor information for over 500 pre-defined glycan compositions, refining for only high-quality precursors, and exporting the relative intensities of each high-quality glycan composition detection. This approach drastically reduces data analysis speed to less than 10 seconds for each sample.