SelectScience
Join Free
Accelerating Science
NewsLife Sciences

Top Methods for Glycobiology Analysis

SelectScience

Discover the top methods for the analysis of glycans, glycopeptides and other glycoforms

15 Feb 2016
Lois Manton-O'Byrne, PhD
Executive Editor

Editorial article

Discover the top methods for the analysis of glycans, glycopeptides and other glycoforms

1. Glycoform Profiling from Therapeutic Antibodies at the Protein, Peptide and Cleaved Glycan Level Using Mass Spectrometry

The glycans (oligosaccharides) attached to proteins play a crucial role in many recognition, signaling, and adhesion events within and between cells. Understanding exactly how the carbohydrates in therapeutic proteins modulate their functions in patients is key to the development of an optimal glycan profile, emphasizing the importance of performing glycan analysis. This application note summarizes the glycoform profiling data from a therapeutic antibody at three different levels: intact protein, peptide (glycopeptides) and cleaved glycans using mass spectrometry (MS).

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

Glycans are widely distributed in biological systems in ‘free state’ and conjugated forms such as glycoproteins, glycolipids, and proteoglycans. This application note demonstrates the development of novel HPLC and UHPLC columns for high resolution separation and structural characterization of native and fluorescently labeled N-glycans released from various proteins, including antibodies.

3. Describing N-glycan Structures in mAb Analysis

In this seminar, Dr Oscar Potter, R&D Scientist at Agilent Technologies, discusses N-glycan structures, names and symbol nomenclature for biopharma scientists. It’s well known that monoclonal antibodies (mAbs) contain N-glycan structures, so it is important to understand and describe these structures when analyzing mAbs.

4. Analysis of Glycopeptide Glycoforms in Monoclonal Antibody Tryptic Digest Using a UPLC HILIC Column

This application note presents a UPLC HILIC/TUV/MS method for the separation of glycopeptides that is complementary to HILIC/FLR separation of N-linked glycans released from the protein. The method does not require glycan release and labeling, as such complexity of sample preparation is reduced, making it useful in the development and quality control of new protein-based therapies.

5. Advance Your Glycan Mapping Through Higher Speed and Resolution

In the race to discover the next promising biotherapeutic or develop a reliable biosimilar, you cannot compromise on analytical accuracy and efficiency. Reducing process development time, having the ability to make procedural changes quickly, and minimizing variability in your analyses are just a few of the challenges you face every day. Learn how to achieve new levels of resolution for glycan mapping with Agilent AdvanceBio Glycan Mapping columns in this application note.

6. Processing Human Serum for Rapid and Reproducible N-Glycan Mass Profiling

This application note describes a high throughput sample preparation procedure for global glycan mass profiling. Microwave-assisted enzymatic N-glycan release followed by an automated solid phase extraction (SPE) purification procedure utilizing the Gilson GX-274 ASPEC™ System were employed to improve sample throughput and stabilize sample processing.

7. Comprehensive Study of O-linked Glycopeptides of Erythropoeitin

Protein glycosylation can effectively be analyzed on the glycopeptide level where site-specific modifications remain preserved. This application note introduces a new method allowing for automatic identification of unknown glycopeptides in one step without prior release of the glycans.

Find out more about glycan research by visiting our Biopharmaceutical Special Feature for the latest news, methods and products!

Related products

Request Quote for All Products

Links

SelectScienceCompany website

Tags

UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.AntibodiesAntibodies are used in techniques such as confocal and fluorescence microscopy, flow cytometry, ELISA, ELISPOT, immunohistochemistry, western blotting and immunopreciptation. Select specific antigen reactivity, high specific affinity, low non-specific binding, monoclonal or polyclonal, primary or secondary antibodies and associated conjugates such as an enzyme or dye for visualization.GlycobiologyGlycobiology encompasses research into the biological functions, synthesis and structures of glycans, including glycoproteins, glycolipids, proteoglycans and free oligosaccharides. Mass spectrometers and their software are commonly used in the study of glycobiology, along with structure predicting tools, chromatography and NMR.MicroplatesMicroplates are multi-well plates used to increase the throughput of biological investigations. The number of wells microplates contain can range from 6 to 3243 wells, with the 96-well format being the most commonly used. Microplates can come tailored for a range of applications including cell culture, PCR, filtration, storage, non-binding surface, protein crystallization, as well as pre-coated, deep well and normal Standard microplates. Additionally, options for microplate colors include clear , black, white or black with clear bottom and white with clear bottom for absorbance microplate reader applications. Find the best microplates for your lab in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.PCR and Thermal CyclingPolymerase chain reaction (PCR) kits and thermal cyclers are used for the in vitro amplification of DNA permitting subsequent analysis and experimental procedures. Explore a range of high-quality polymerase, primers and nucleotides or simplify your workflow with a PCR mastermix. Find reverse transcription PCR (RT-PCR) and cDNA synthesis kits for RNA products and libraries. Quantitatively measure the amplification of DNA with real-time PCR (qPCR) and droplet digital PCR (ddPCR) kits and systems, and discover automated PCR setup solutions to increase throughput. Alternative DNA amplification methods also include recombinase polymerase amplification (RPA) kits. Find the best PCR kits and thermal cyclers and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Microplate Readers / DetectorsMicroplate readers are used to automate the detection and analysis of labeled or label-free components in microplates during assays or live-cell monitoring. Microplate readers are generally distinguished by their mode of detection. Types include absorbance, luminescence, fluorescence intensity, fluorescence polarization, TRF / FRET and multimode microplate readers. Microplate readers deliver a high throughput of samples by reading multiple wells simultaneously, with the 96-well format the most commonly used. As a result, microplate readers are often used in the drug discovery, bioassays, research and pharmaceutical industries for screening applications. Microplate loading can also be automated, with robotic microplate stackers to increase throughput. Find the best microplate readers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.GlycansMonoclonal AntibodiesMonoclonal antibodies (mAbs) are lab-made molecules that can bind to specific antigens on cells, making them highly effective in treating cancers, autoimmune diseases, and infections. These targeted therapies are transforming the treatment landscape for many conditions. Browse our peer-reviewed product directory to find monoclonal antibodies for research and clinical applications; compare products, check reviews, and get pricing directly from manufacturers.BiopharmaceuticalsBiopharmaceuticals are proteins and other compounds (such as nucleic acids) produced by living organisms that have uses as therapeutics or for in vivo diagnostics. The most well known example of a biopharmaceutical product, and the first to be approved for therapeutic use, was recombinant human insulin.PeptidesPeptides are short chains of amino acids that play key roles in biological processes, including signaling, immune responses, and enzyme regulation. Peptide research is critical in drug discovery, diagnostics, and vaccine development. Browse our peer-reviewed product directory to find the best peptides and peptide-related products, compare options, check reviews, and get pricing directly from manufacturers.BiotherapeuticsBiotherapeutics are proteins and other compounds (such as nucleic acids) produced by living organisms that have uses as therapeutics or in <i>in vivo</i> diagnostics. The most well-known example of a biotherapeutic product, and the first to be approved for therapeutic use, was recombinant human insulin.Protein StructureProtein structure refers to the three-dimensional arrangement of amino acids in proteins, determining their function. Understanding protein structure is key in drug discovery, enzymology, and molecular biology. Explore protein structure analysis tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Expression Profiling

Related articles