High Throughput Post-Labeling Cleanup of Biopharmaceutical Glycans

29 Nov 2011
Roger Wayman
Administrator / Office Personnel

Product news

Ludger has developed a fast, high-throughput system for post-labeling cleanup of glycan samples. The LudgerClean T1 cartridge system is designed for drug glycoprofiling analysts who want the reliability of S-cartridges but also want to process larger numbers of samples easily and get the results more quickly. The cartridges work with a Ludger-Velocity™ SPE vacuum manifold.

HPLC analysis of fluorescently labeled oligosaccharides is a widely used tool for biopharmaceutical glycoprofiling. In this process, glycans are released from the therapeutic glycoprotein then tagged with a fluorescent dye such as 2-AB (2-aminobenzamide) or 2-AA (2-aminobenzoic acid) by reductive amination prior to chromatography. Excess labeling reagents such as dye and reducing agent interfere with the HPLC analysis - so need to be removed without disturbing the relative proportions of the various glycan species.

One method for achieving post-labeling cleanup is the use of LudgerClean S-cartridges. These employ a hydrophilic membrane to separate the labeled glycans from non-glycan contaminants and are routinely used in the biopharma industry during glycoprofiling QC for lot release of FDA and EMA approved biopharmaceuticals. They are also the method of choice in our laboratories for detailed characterization of drug glycosylation patterns for regulatory submissions. S-cartridges work well but they are slow and not suited for high throughput (HT) work. For this reason Ludger has developed the LudgerClean T1 cartridge system.

Links

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.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.Solid-Phase ExtractionSolid-phase extraction (SPE) is used for clean-up, extraction and concentration of semi-volatile or non-volatile analytes from complex mixtures including blood, urine and food samples. Multiple formats are available for conducting SPE, including prepacked SPE cartridges, disks and microplates, as well as SPE sorbent powders for manual packing. SPE systems can be used to automate the process and extract multiple samples at once. Solid-phase microextraction (SPME) and supported liquid extraction (SLE) products are also available in the same formats. Find the best SPE, SLE and SPME equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Biopharmaceutical AdvancesBiopharmaceutical advances follow the development of pharmaceuticals derived from biotechnology, also known as biotechnology medicines. Biopharmaceuticals may be produced from cell lines, plants, or microbial cells. Important considerations of biopharmaceutical use include application, cost, production process and purification.GlycansPost-LabelingQuality ControlQuality control is needed in all production processes. Quality control is a monitoring procedure or set of procedures that are put in place to ensure that a manufactured product adheres to a defined set of quality criteria. Labeling