ResourceSeparations

Advancing Host Cell Protein Analyses Through the Combined Use of Microscale 2D RP/RP with CSH C<sub>18</sub> and Ion Mobility Enabled MS Detection

26 Feb 2015

In this application note, recent advances in microscale peptide chromatography are applied to HCP analysis. The novel charge-surface-modified C18 material, known as CSH C18, which can produce high peptide peak capacities even at high mass loads, is used in the format of a 15K psi capable 300 μm I.D. column along with an ACQUITY UPLC M-Class based 2D-RP/RP system and SYNAPT HDMSE detection to provide a robust analytical workflow for the identification of low concentrations of HCP impurities at single digit PPM levels (parts per million, or ng/mg).

ACQUITY UPLC M-Class System

Waters

The analysis you’ve always desired for nano- to microscale separations The ACQUITY UPLC M-Class System delivers the most robust, reliable, and highest quality nano- to microscale UPLC separations to every laboratory performing LC/MS analyses. With unprecedented usability, the ACQUITY UPLC M-Class System enables laboratories to succeed in analyzing compounds that are severely limited in amount or availability, even amid a complex matrix over several orders of magnitude. Confidently achieve your laboratory goals with high-efficiency ACQUITY UPLC M-Class Columns ranging from 75 µm to 1 mm I.D. by using complementary mass spectrometry to support our full range of column I.D.’s, or run 300 µm to 1 mm I.D. columns with optical detection.The system also offers a range of flexible configurations for HDX technology, UV detection, and 2D analysis.ACQUITY UPLC M-Class System Features: Highest sensitivity - UPLC separations yield increased analyte concentration for higher UV and MS signal responses Maximum chromatographic resolution - New 15k psi-capable ACQUITY UPLC M-Class Columns deliver sub-2-µm particle performance Faster nano- and microscale separations - Increased operating pressures yield maximum flexibility for higher throughput analyses Higher resolution with longer columns - Increased pressure enables more separation power Confidence in sample analysis - New non-reactive wetted surfaces throughout the fluidic path minimize absorptive losses Increased solvent compataibility - Improved range of solvent blends precisely metered for gradient fidelity Increased sample capacity - Sample organizer compatibility allows you to store a wide variety of plates and vials in the system for longer, unattended operation UV compatibility - UV flow cells compatible with 300 µm to 1.0 mm I.D. columns to augment your UPLC/MS analysis Scalability - With flow rates from 200 nl/min to 100 µL/min, achieve the best chromatographic performance, from 75 µm to 1.0 mm I.D. columns covering a 170-fold range of column sizes Increased flexiblity - More value configurations enable a variety of applications, icnreased throughput, and maximized peak capacity

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XBridge® and ACQUITY UPLC® Oligonucleotide C18 Columns

Waters

XBridge® and ACQUITY UPLC® Oligonucleotide C18 Columns for Synthetic Oligonucleotide Analysis and PurificationIon-pair, reversed-phase separation of detritylated oligonucleotides on sub-3 micron particles yields exceptional component resolution.UPLC ® technology increases sample throughput while maintaining necessary oligonucleotide component resolution.LC/UV and LC/MS methods for enhanced quantitative and qualitative oligonucleotides analyses.Various XBridge™ Oligonucleotide C 18 column dimensions address lab scale purification needs.2nd generation hybrid-silica BEH Technology™ particles deliver exceptional column life.Use of synthetic oligonucleotides for research, diagnostic, and therapeutic applications continues to increase. The availability of 2.5 μm HPLC or 1.7 μm UPLC ® technology particles in several column dimensions provides flexibility to meet various LC/UV or LC/MS application needs. Waters' stringent manufacturing and quality control procedures help ensure consistent batch-to-batch and column-to-column performance.

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SYNAPT G2-Si MS

Waters

Transform your lab's discovery capability with the SYNAPT G2-Si MS. Information. Informatics. Impact. SYNAPT enables extensive characterization of complex mixtures and molecules with uncompromising qualitative and quantitative performance, streamlined workflows and unparalleled platform versatility. With the SYNAPT G2-Si you get ultimate UPLC/MS/MS performance, powerful data independent & data dependant solutions, CID and ETD fragmentation capabilities, and a wide range of experimental options.

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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.Ion ChromatographyIon chromatography, also known as ion exchange chromatography, is a high-performance liquid chromatographic technique used for the separation and identification of ions or polar molecules in a sample, including proteins, nucleotides and amino acids. Equipment includes ion exchange columns, ion exclusion columns, ion chromatography systems, pumps, and detectors. Find the best ion chromatography equipment 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.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.Ion Mobility Spectrometry