Sepax Proteomix Brochure - Winlab.com.au

More documents

Recommendations

Info

Particle Size Impact on the SeparationThe smaller the particle size, the better separation efficiencyand higher separation speed. Proteomix ion-exchangeresins have a wide range of the particle size selection from1.0 to 10 µm for analytical and semi-preparative separationapplications. Figure 9 is the separation of three proteins byfour weak cation exchange resins with the particle size of1.7, 3, 5, and 10 µm, separately. It is clearly seen that thesmaller particle generates higher efficiency and resolution.One of the impurities in lysozyme labeled by Peak N isbarely separated with10 µm WCX resin. For 5 µm WCXresin, it is almost a baseline separation. When the particlesize is decreased to 3 µm, it becomes a baselineseparation. With the particle size further decreased to 1.7µm, the impurity is well separated.Fig. 9. Separation of a protein mixture by Proteomix WCX-NPcolumns with various particle sizes.Charge Dependent SeparationFigure 10 is a typical test chromatogram for a ProteomixSCX-NP column (3 µm). Extremely high efficiencyseparation was achieved for 5 proteins. Those proteinswere eluted based on their charge status (pI) from low tohigh: ovalbumin, ribonuclease A, cytochrome C, aprotinin,and lysozyme. At the separation conditions (pH 6.0),ovalbumin is negatively charged and eluted at the voidvolume, indicating no non-specific bindings.Fig. 10. Separation of a protein mixture with various pI by aProteomix SCX column.ProteinspI1. Ovalbumin 4.62. Ribonuclease A 8.73. Cytochrome C 9.64. Aprotinin 10.05. Lysozyme 11.012Proteomix WCX-NP(4.6x50mm)mAU12534043301.7 µm3 µm5 µm10 µmPeak N20100 2 4 6 8 10 12 14 16 18 20 22Min2 4 6 8 10 12minColumn: Proteomix WCX-NP (4.6x50 mm)Mobile phase: A, 10 mM phosphate, pH 6.0B, A + 1.0 M NaClGradient: 0-100%B in 20 minFlow rate: 1.0 mL/min, 0.75 mL/min for WCX-NP1.7Sample: 1) Cytochrome C, 2) Ribonuclese A,3) LysozymeInjection: 5 µL (1 mg/mL for each protein)Temperature: 25 o CDetection: UV 280 nmColumn: Proteomix SCX-NP3 (3 µm, 4.6x50 mm)Mobile phase: A, 10 mM phosphate, pH 6.0B, A + 1.0 M NaClGradient: 0-100%B in 15 minFlow rate: 0.5 mL/minInjection: 5 µL (1 mg/mL for each protein)Temperature: 25 o CDetection: UV 280 nmwww.sepax-tech.com 5 1-877-SEPAX-US
High Lot-to-Lot ReproducibilityWith well-controlled resin production and the surfacechemistry, the manufacturing of the Proteomix ionexchangersis highly reproducible. The typical variation ofthe retention time is less than 6% from batch to batch. Oneexample is shown in Figure 11 for the production of threelots of 3 µm Proteomix SAX-NP resins.Fig. 11. The reproducibility of three lots of Proteomix SAX resins.Proteomix SAX-NP3Lot ALot BLot COvalbuminBSAPorous Protomix Ion-ExchangerProteomix SCX-POR, WCX-POR, SAX-POR, and WAX-POR resins are based on porous PS/DVB resins that arecoated with a proprietary hydrophilic layer andfunctionalized with a uniform ion-exchange layer. Thesephases have three unique features. First, the nanometerthick hydrophilic layer completely eliminates the non-specificinteractions with biological analytes. Second, Sepax’sproprietary technology synthesizes a uniform and denselypacked layer of ion-exchange functional groups. Third,porous beads provide high surface area, leading to highseparation capacity. Such uniquely designed ProteomixSCX-POR, WCX-POR, SAX-POR, and WAX-POR phasesoffer high capacity and reasonably high resolutionseparations for proteins, oligonucleotides and peptides.Due to the porosity, Proteomix porous resins have lessefficiency and lower resolution than Proteomix non-porousresins. Proteomix SCX-POR, WCX-POR, SAX-POR, andWAX-POR phases are well suited for semi-preparative andpreparative separation and purification of biomolecules.Porous Proteomix SCX, WCX, SAX, and WAX columns aremanufactured and tested under the strictest specifications,resulting in high column-to-column and lot-to-lotreproducibility.0 2 4 6 8 10 12 14minColumn Dimension AvailabilityColumn: Proteomix SAX-NP3 (3 µm, 4.6x50 mm)Mobile phase: A, 20 mM Tris, pH 8.0B, A + 0.3 M NaClGradient: 0-100%B in 15 minFlow rate: 0.5 mL/minSample: 1) Ovalbumin (10 mg/mL), 2) BSA (5mg/mL)Injection: 10 µLTemperature: 25 o CDetection: UV 214 nmMono-dispersedProteomix ionexchangeparticlesenable uniform columnpacking bed, whichimproves separationefficiency.The column dimensions of the Proteomix SCX, WCX, SAX,and WAX products are 0.75, 2.1, 3.0, 4.6, 7.8, 10, and 21.2mm I.D., and 2, 3, 5, 10, 15, 25, and 30 cm length. Sepaxalso offers custom-made columns.Features Uniform polymer beads as the packing support Proprietary surface chemistry specially designed forelimination of non-specific bindings Unprecedented separation efficiency, selectivity andresolving power Complete selection for analytical, semi-preparative andpreparative separations Wide particle size selection High stability High recovery Well suited for UPLC systemwww.sepax-tech.com 6 1-877-SEPAX-US
Page 3 and 4: Proteomix ® Ion-exchange PhasesGen
Page 5: capacities. Such innovatively desig
Page 9 and 10: Separation of Carbohydrates and Gly
Page 11 and 12: Separation and Analysis of Cell Lys
Page 13 and 14: PhaseID x Length((mm)P/N Price Phas
Page 15: Proteomix WCX-POR ColumnsPhaseID x

Sepax Proteomix Brochure - Winlab.com.au

Create successful ePaper yourself

Delete template?

Save as template?