Posters

Mass Spectrometry-based High-throughput Profiling of Porcine Notochordal-Cell Matrix N-glycans Using GlycanExplorer^™ Software

N-glycans play key roles in important biological processes such as protein folding and tracking, cell-cell interactions, and signal transduction [1]. This study analyzes the N-glycome of Porcine notochordal cell-rich nucleus pulposus matrix (NCM) [2].

Goswami, R., Joyce, K., Günay, B., Hing, M.N.T., Walsh, C., Schmitz, T., Tryfonidou, M., Ito, K., Kolli, K., Pandit, A., Saldova, R., Apte, A.

71^st ASMS Conference on Mass Spectrometry and Allied Topics (2023), Houston, TX

Comprehensive N-glycan Profiling of USP Monoclonal Antibody Reference Standard Using GlycanExplorer^™ Software

Glycosylation is the most abundant and complex post-translational modification in proteins. It is considered a critical quality attribute of therapeutic glycoproteins such as monoclonal antibodies (mAbs) because the glycans alter critical properties of a protein such as binding, stability, folding, immunogenicity, and turnover [1]. Comprehensive characterization of glycans is necessary to understand the therapeutic “behind the scene” properties of glycoproteins. The United States Pharmacopeia (USP) has released three mAb reference standards, (i.e., USP mAb 001, USP mAb 002, and USP mAb 003) that can work as controls to help establish an efficient glycan characterization protocol [2].

Apte, A., Goswami, R., Kolli, K.

71^stASMS Conference on Mass Spectrometry and Allied Topics (2023), Houston, TX

Introducing a Novel Software Containing an Experimental High-Resolution Mass Spectral Library of N-Glycans and Automated Glycan Mapping Using UHPLC-Orbitrap-MS Analysis

The therapeutic use of monoclonal antibodies (mAbs) and fusion proteins has revolutionized the treatment of several diseases. Glycosylation of therapeutics must be fully characterized during biotherapeutic development and manufacture; with tandem-MS based methods commonly used for glycan characterization. However, the MS-based approach can be challenging, primarily due to uncertainty in software-derived glycan identifications.We have developed novel software for glycan identification for the biopharmaceutical industry containing an integrated MS2 spectral library. The library currently contains spectra for 42 unique fluorescently labeled N-glycans from a wide range of glycoprotein sources with data acquired using Orbitrap-based MS instruments. The novel software workflow features LC-MS data processing, glycan mapping and a spectra pattern matching algorithm for rapid and accurate identification of glycan structures.

Goswami, R., Kolli, K., Sharath, A. K., Apte, A.

69^th ASMS Conference on Mass Spectrometry and Allied Topics (2021), Philadelphia, PA