Size Exclusion Chromatography (SEC)-based Oligosaccharides Analysis Service
Oligosaccharides are complex carbohydrates playing pivotal roles in diverse biological processes, including cell recognition, immune response modulation, and protein folding. The precise analysis of their size distribution and quantitative profiling is critical in fields ranging from enzyme kinetics to biopharmaceutical characterization and food science. CD BioGlyco offers an advanced size exclusion chromatography (SEC)-based oligosaccharides analysis service, providing a robust and highly accurate solution for the qualitative and quantitative profiling of oligosaccharide mixtures. This service is designed to support clients in understanding the structural and functional attributes of these vital biomolecules.
High-performance size exclusion chromatography (HPSEC) serves as the core analytical platform for our oligosaccharide characterization service. HPSEC separates molecules based on their hydrodynamic volume (size) in solution through differential pore access within the stationary phase. Smaller molecules permeate deeper into the porous matrix, resulting in longer retention times, while larger molecules experience restricted pore entry and elute more rapidly.
Our SEC-based oligosaccharides analysis service delivers comprehensive characterization for diverse oligosaccharide mixtures.
We provide detailed chromatographic profiles that illustrate the size distribution of oligosaccharides within a sample, essential for understanding the complexity of your mixture.
Our robust methodology allows for the precise quantitative profiling of oligosaccharide mixtures, enabling the determination of relative and absolute amounts of different size fractions. This is critical for applications like monitoring enzymatic hydrolysis progress.
The service is highly effective for characterizing the products of enzyme-catalyzed hydrolysis of various carbohydrate substrates, such as plant biomass materials. This allows for accurate assessment of enzyme activity and product distribution.
Our expertise and optimized methods allow for the analysis of a broad spectrum of oligosaccharides, from disaccharides to complex higher oligomers.
We initiate each project with a detailed consultation to understand your specific research objectives, sample types, and desired analytical outcomes. This ensures the design of a tailored SEC method.
Samples are carefully prepared, which may involve dilution, filtration, or desalting to ensure compatibility with the SEC system. Each sample undergoes preliminary qualification to assess its integrity and concentration.
Our expert team develops and optimizes the HPSEC method for your specific oligosaccharide mixture. This involves selecting appropriate SEC columns, optimizing mobile phase composition (e.g., ionic strength, pH), flow rate, and temperature to achieve optimal resolution and separation of oligosaccharides by size.
Prepared samples are injected into the HPSEC system. Oligosaccharides are separated based on their hydrodynamic volume as they pass through the column, with their elution monitored in real-time by a refractive index detector.
Elution profiles are meticulously recorded. The quantitative analysis involves dividing HPSEC elution profiles into fixed time intervals and utilizing the linear refractive index response (area under the curve) of defined standard compounds for calibration.
The processed data are then used to generate quantitative profiles of the oligosaccharide mixture. A comprehensive report is provided, including elution profiles, peak integration, quantitative data, and expert interpretation of the results.
Journal: Antibody Engineering
DOI: 10.5772/intechopen.73321
Published: 2018
Results: This study details the application of analytical SEC for separating and characterizing monoclonal antibodies (mAbs). SEC separates molecules based on their size/hydrodynamic radius in solution, making it ideal for resolving mAb monomers (~150 kDa) from critical impurities like high molecular weight (HMW) aggregates (dimers, trimers) and low molecular weight (LMW) fragments. Achieving >99% purity is essential as aggregates can cause immunogenic responses. The author emphasizes key factors for successful SEC analysis: selecting columns with appropriate pore size (e.g., 25-30 nm) and optimized surface chemistry (e.g., diol coating) to minimize unwanted secondary interactions, and optimizing mobile phase conditions (buffer, pH, ionic strength, potential additives like IPA). It also discusses the SEC's role in forced degradation studies, MS-compatible methods, orthogonal techniques for detecting heterogeneity, method transfer to UHPLC, and troubleshooting tips. SEC is presented as a critical QC tool for mAb purity assessment.
Our SEC-based oligosaccharide analysis service provides precise characterization of oligosaccharide size distribution and purity, laying the essential groundwork for functional glycan engineering. Building on this analytical foundation, we offer advanced cell surface engineering solutions that strategically modify cellular glycans for targeted applications:
CD BioGlyco is your premier partner for advanced oligosaccharide research, offering integrated SEC-based analysis services alongside Custom oligosaccharide synthesis. Driven by expert experience and cutting-edge technology, our solutions deliver unparalleled precision in quantitative profiling and tailored molecular design, accelerating scientific discovery and development. today to discuss your project needs, obtain a detailed quote, or learn how our combined synthesis-to-analysis capabilities advance your research.
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