SimGlycan: Facilitating Automated Evaluation of Glycoengineered Therapeutic Glycoproteins using MS Methods

Monoclonal antibody (mAb) based therapeutics are the fastest growing class of human pharmaceuticals [1]. Variation in IgG Fc glycoforms affects the safety, clinical efficacy, and effector functions of therapeutic antibodies including antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) e.g., the terminal galactose enhances CDC activity, but fucose significantly decreases ADCC [2].

Meitei, N.S., Goswami, R.

SFG 2018 Annual Meeting, New Orleans, Louisiana

Structural Identification, and Quantitation of Glycans Labeled with Dual Modifications Strategies from Complex Mixtures using LC-MS Workflows with SimGlycan^® Software

Glycans play a critical role in the therapeutic efficacy and toxicity of monoclonal antibody proteins. Recently developed integrated chemical strategy, Dual Reactions for Analytical Glycomics (DRAG), enables quantitative comparison between neutral and sialylated glycans simultaneously using mass spectrometry methods [1].

Meitei, N.S., Li, H., Saha, S., Goswami, R., Apte, A., Lee, R.S.

66^th ASMS Conference on Mass Spectrometry and Allied Topics (2018),
San Diego, CA

Identification and Subsequent Relative Quantitation of Stable Isotope Labeled Released N-Glycans by LC-, ESI-, Tandem MS with SimGlycan^® Software

The Monoclonal Antibodies (mAb) are proved indispensable for treatments of various immune-mediated disorders and thus, of high importance to therapeutic industries. Glycans play a critical quality attribute for the therapeutic monoclonal antibodies, where terminal sugars of the Fc glycans are responsible for determining their efficacy and safety. Glycomics analysis involves correct identification of the glycans as well as their subsequent quantitation in the samples.

Meitei, N.S., Goswami, R., Jonggyu, P.

Mass Spectrometry & Proteomics 2018, Osaka, Japan

Automated Identification and Quantitation of Stable Isotope Labeled Released N-glycans by LC-MS with SimGlycan^® Software

Glycans play a critical quality attribute for the therapeutic monoclonal antibodies, where terminal sugars of the Fc glycans are responsible for determining their efficacy and safety. One of the emerging trends of glycomics research include quantitative and qualitative analysis of glycans labeled with tandem mass tags or isotope labels.

Automated Identification and Subsequent Relative Quantitation of Glycans Using Stable Isotope Labels by MS with SimGlycan^® Software

Recent trends in glycomics research show rapid growth of innovation related to accurate mass spectrometry based quantitative analysis of glycans. Stable isotope labels facilitate accurate identification and subsequent quantification of glycans using LC-MS or ESI-MS workflows.

Meitei, N.S.

SFG 2017 Annual Meeting, Portland, Oregon, Hilton Portland Downtown

Automated and Rapid Characterization of Low-Molecular-Weight Heparin Oligosaccharides Using TripleTOF 5600 Mass Spectrometer with SimGlycan Software

Low molecular weight heparins (LWMH) - Enoxaparin, Parnaparin, Certoparin, Deltaparin, Tinzaparin, etc. - have been reported to be the key drugs and are as safe and effective as Unfractionated Heparin (UFH) [1,4] which further necessitates in-depth study of new regulatory drugs. Despite its medical and biological importance, LWMHs are relatively uncharacterized in terms of its chemical structures due to its structural complexity.

SimGlycan®: Informatics Support for Automated Characterization of Low-Molecular-Weight Heparin Oligosaccharides Using MS2 Data from TripleTOF® 5600 Mass Spectrometer

Recent reports suggested that patent protection worth of $80 billion for several biosimilars, including Low molecular weight heparins (LWMH) - Enoxaparin, Parnaparin, Certoparin, Deltaparin, Tinzaparin, etc. - has been expired and abbreviated new drug applications have been approved by different regulatory bodies, namely, the Food and Drug Administration (FDA), USA; Central Drugs Standard Control Organization (CDSCO), India, etc.

Speeding up the High Throughput Searches for Glycan Analysis

A typical LC-MS glycomic data processing includes peak detection, deconvolution of peaks of co-eluting isomers, molecular feature extraction, and selection of correct precursor m/z values from the isotope cluster for MS/MS data analysis.

Development of Bioinformatics support for Quantitative Glycomics using Tandem Mass Tags

AminoxyTMT reagents enable relative quantitation of glycans in a MS/MS spectrum. However, there are no bioinformatics tools to automate the qualitative/ quantitative analysis.

Profiling the Distribution of N-Glycosylation in Therapeutic Antibodies using the QTRAP^® 6500 System

Immunoglobulin G molecules have become attractive as targeted therapeutic proteins, due to their high specificity and long circulation time. Glycosylation patterns determine the stability and bio-disposition of these recombinant protein drugs in vivo, as well as the efficacy, folding, binding affinity, specificity and pharmacokinetic properties.

SimGlycan: Informatics tool for creating glycan templates to enable high throughput glycan identification from Liquid Chromatogram - tandem mass spectrometry data

Protein glycosylation is one of the well-known post-translational modifications, and in biological processes in living organisms it plays an important role. Monitoring changes in the amount and structure of sugar chains makes it possible to monitor sugar chain modification of proteins and various diseases.

Development of Bioinformatics Support for High Throughput Isomeric Separation and the Structural Identification of Glycans by LC-MS

The separation of glycans by chromatography prior to MS analysis can reduce sample complexity, minimize ion suppression, and increase dynamic range and separation of structural isomers.

Informatics Support for Isomeric Separation and the Structural Identification of Labeled N-Glycans from Proteins

Develop a software to facilitate LC-MS and MS/MS glycan data that have been acquired for separation and identification of various complex N-linked glycans from proteins using a novel high resolution mixed-mode column and an Orbitrap Fusion Tribrid mass spectrometer.

Automating Mass Spectrometry-Based Quantitative Glycomics using Tandem Mass Tag (TMT) Reagents with SimGlycan

One of the emerging trends in glycomics research is the innovation related to accurate MS based quantitative analysis of glycans1-4. Recently, we have introduced aminoxyTMT Reagents, which enable efficient relative quantitation of carbohydrates, improved labeled-glycan ionization efficiency and increased analytical throughput.

Automated Glycan Structural Isomer Differentiation Using Bioinformatics Tool

Mass spectrometry (MS) has emerged as a powerful tool for the structural elucidation of glycans. The use of permethylation in combination with multistage fragmentation (MSⁿ) is a critical step in glycan structural characterization.

Integration of SimGlycan into Waters Glycan/Glycopeptide Analysis Workflow

Translating New Technologies in Improved Workflows for Biotherapeutic Analysis.

A Unique Workflow for Glycoprotein Characterization from Sample Preparation to MS/MS Spectral Interpretation

Protein glycosylation is a complex dynamic post-translational modification, which is used by an organism to regulate a number of important protein function.

SimGlycan™ Software*: A New Predictive Carbohydrate Analysis Tool for MS/MS Data Automated Data Interpretation for Glycan Characterization

The analysis of carbohydrates is of great importance in modern biochemistry and is often tackled using various analytical techniques, including mass spectrometry.

Novel Glycan Column Technology for the LC-MS Analysis of Labeled and Native N-Glycans Released from Proteins and Antibodies

Because glycosylation is critical to the efficacy of antibody therapeutics, the FDA requires that a consistent human-type glycosylation be maintained for recombinant monoclonal antibodies (mAb), irrespective of the system in which they are produced.