AL
and lower baseline noise, were also exhibited by Shodex.
MAbPac RP columns from Thermo Fisher Scientific are designed for top-down pro- teomics. The particle is a 4-μm-diameter polymer with a 1500- Å pore, wide enough for antibodies and antibody–drug conjugates. The column packing can withstand 0.8 M NaOH at 80 °C, which is used as a regenerant solution to clean protein residue from the stationary phase. The polymeric bead is com- patible with all common mobile phases used in LC/MS of proteins.
Tosoh (Tokyo, Japan) featured Toyopearl AF- rProtein A HC-650F resin for the capture and release of antibodies for sample prep, process research and production-scale chromatogra- phy. Protein A can cost as much as $17,000 per liter. Tosoh notes that the high-capacity resin, indicated by the “HC” in the column’s name, often provides the lowest cost per gram of purified target.
Welch Materials Inc. (Austin, Texas) introduced Boltimate core-shell HPLC columns. Packed with 2.7-diameter particles with a 0.5-μm porous shell encasing a 1.7-μm solid silica core, the columns have a sub-2-μm column efficiency (~200,000 plates per meter) while avoiding the ultrahigh-pressure region of UHPLC. A direct-connect cartridge-style guard column is also offered for the Boltimate. For UHPLC, Welch offers columns packed with totally porous particles with nominal particle sizes of 1.8, 3.0 and 5 μm. Surface chemistries include C18, phenylhexyl, perfluorophenyl and hydrophobic interaction liquid chromatogra- phy (HILIC). Inline filters protect the columns from particulate contamination, often cited as a cause of pump and injector seal wear.
Waters extended the CORTECS solid-core LC column line by adding CORTECS C8 and CORTECS Phenyl surface chemistry bonded to 1.6- or 2.7-μm particles. The C8 product fills the gap created by some compendial methods that specify C8 surface chemistry. The phenyl phase provides useful selectivity for aromatic analytes.
Osaka Soda Co. Ltd. (Tokyo, Japan) (formerly Daiso Co.) markets silica-based LC column packings with nominal pore sizes ranging from 60 to 120, 200, 300, 1000 and 2000 Å.
Particle sizes start at 1.7 μm and extend up to 40/60 μm for preparative separations. A wide selection of bonded-phase surface chemis- tries is also available.
N-glycan kit Precise control is needed during the production of biotherapeutics, as the function of many pro- teins depends on the structure and location of glycosylation. Generally, the glycol groups are cleaved from the protein, fluorescently labeled and measured by LC. MS is also used.
Regeneron Pharmaceuticals (Tarrytown, N.Y.) compared its existing biotherapeutic produc- tion process with one using the new GlycoWorks RapiFluor-MS Kit from Waters. Regeneron reports the current workflow has turnaround times of tens of hours; the RapiFluor kit reduces turnaround time to about 45 minutes. New cleavage and derivatization protocols greatly accelerate the assay. Regeneron expects to use the RapiFluor along with Waters UPLC with QDa detection in the QA workflow for glycoprotein therapeutics. The RapiFluor-MS kit has reportedly been adopted by the top 15-biopharmaceutical firms.
Ion chromatography
The Thermo Fisher Scientific Dionex Integrion HPIC (high-pressure ion chromatograph) in- cludes columns packed with 4-μm particles, which improves chromatographic resolution and speed (see Figure 6). A new electrochemi- cal detector is optional, previously available only with the Dionex ICS-5000. The plumbing of the Integrion has been simplified to provide reliable, leak-free operation with Dionex Viper fittings. A thermally regulated detector com- partment is included as well. The Chromeleon Chromatography Data System (CDS) controls the instrument and provides data reduction and report generation. Whole instrument smart monitoring detects deviations before they be- come problematic.
A new compact ion chromatograph, the Dionex Aquion, replaces the Dionex ICS-1100 platform. The new platform offers electrolytic suppres- sion to provide high-sensitivity detection and convenience. The IC will probably be used as a dedicated analyzer that can run the same assay for years. A column heater and vacuum degasser are optional.
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