Sepax Proteomix Brochure - Winlab.com.au

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Non-Porous Proteomix Ion-exchangers– Technology AdvancementsHigh Capacity of Proteomix Non-Porous ResinsIt is well known that the non-porous resin maximizes themass transfer and minimizes the lateral diffusion, resultingin high speed and high efficiency separations. However, dueto the low surface area, the separation capacity is low. Thelow capacity issue creates a great challenge to the nonporousresins for many applications. The Proteomix nonporousion-exchange resins developed by Sepax proprietarytechnologies have a breakthrough on the improvement oftheir capacities. This new technology enables the capacityof the non-porous resins increased to the level comparableto that of the porous resins. Figure 3 shows examples ofdynamic binding capacities of the 3 µm non-porousProteomix SAX and WAX resins: 35 and 26 mg/mL,respectively. Figure 4 shows an example of the dynamicbinding capacity of 3 µm Proteomix SCX non-porous resin.Its dynamic binding capacity reaches as high as 53.5mg/mL.High dynamic binding capacity leads to high sample loadingwithout deterioration of the separation efficiency and theresolution. Figure 5 shows the elution of cytochrome C withthe loading up to 200 µg for a 1.7 µm Proteomix SCX nonporouscolumn without any deterioration of the separation.AUFig. 4. The dynamic binding capacity test for Proteomix SCX-NP3 resin by flowing lysozyme into the column.mAU150010005000Proteomix SCX-NP3(3µm, 4.6x50 mm)Capacity: 53.5 mg/mL5 10 15 20 25 30minMobile Phase: 10 mM phosphate, pH 6.0Flow Rate: 0.5 mL/minDetection: 280 nmTemperature: 25 o CSample: Lysozyme (3 mg/mL)Fig. 5. Elution profiles of cytochrome C at various loadings.2.01.81.61.41.21.00.8SCX-NP1.7 (1.7µm, 4.6x50mm)200 µg100 µgFig. 3. The dynamic binding capacity tests for Proteomix anionexchangeresins by flowing BSA into the columns.0.60.40.20 4 8 12 16 20Min50 µg(A) Proteomix WAX-NP3(3µm, 4.6x50 mm)0.015 16 17 18 19 20 21 22Minutes(B) Proteomix SAX-NP3(3µm, 4.6x50 mm)Dynamic Binding Capacity• SAX-NP3: 35 mg/mL• WAX-NP3: 26 mg/mLABColumn: Proteomix SCX-NP1.7 (1.7 µm, 4.6x50 mm)Mobile Phase: A, 20 mM PBS, pH 6.0; B, A+1.0 M NaClGradient: 0-70%B (21 min)Flow Rate: 0.35 mL/minDetection: 280 nmTemperature: 25 o CSample: Cytochrome C (20 mg/mL)5 10 15 20 25 30 35minColumn: 4.6x50 mmMobile Phase: 10 mM Tris/HCl, pH 8.0Flow Rate: 0.25 mL/minDetection: 280 nmTemperature: 25 o CSample: BSA (3 mg/mL)High Separation Efficiency, Resolution and SelectivityProteomix SCX-NP, WCX-NP, SAX-NP, and WAX-NPresins have three unique features. First, their nanometerthick hydrophilic layer completely eliminates the non-specificinteractions with the biological analytes. Second, nonporousbeads minimize the biological analytes’ lateraldiffusion and suppress their diffusion into thechromatographic bed. Third, a uniform layer of ionexchangefunctional groups synthesized by Sepax’sproprietary technology greatly improved their ion-exchangewww.sepax-tech.com 3 1-877-SEPAX-US
capacities. Such innovatively designed Proteomix SCX-NP,WCX-NP, SAX-NP, and WAX-NP phases result in thehighest efficiency separations for proteins, oligonucleotides,carbohydrates, and peptides. Figure 6 is a typical testchromatogram for separation of three proteins: ribonucleaseA, cytochrome C, and lysozyme by a 4.6 × 50 mm,Proteomix SCX-NP column (3 µm). The efficiency oflysozyme reaches 100,000 of plates with 5 cm long column.Such a high efficiency separation is unprecedented.Fig. 6. Separation of a protein mixture by a Proteomix SCX-NPcolumn.Fig. 7. The elution profile of lysozyme from its multipleimpurities by a Proteomix SCX-NP column (3 µm, 4.6 × 50 mm).The separation conditions are the same as those in Fig. 6.541/236mAU1006.375111.065Fig. 8. Elution profile of a mixture of ovalbumin and BSA byProteomix SAX-NP5 column.8060407.27323mAUOvalbuminBSA2040002 4 6 8 10 12 14min2000Column: Proteomix SCX-NP3 (3 µm, 4.6x50 mm)Mobile phase: A, 10 mM phosphate, pH 6.0B, A + 1.0 M NaClGradient: 0-70%B in 15 minFlow rate: 0.5 mL/minSample: 1) Cytochrome C, 2) Ribonuclese A,3) LysozymeInjection: 5 µL (1 mg/mL for each protein)Temperature: 25 o CDetection: UV 280 nmThe uniqueness of non-porous Proteomix SCX, WCX, SAX,and WAX phases offers highest resolution and selectivity forprotein separations. Figure 7 shows an elution profile oflysozyme by a Proteomix SCX-NP3 phase (3 µm). Sixpeaks are well resolved with a short column (4.6 × 50 mm).Such a high selectivity and resolving power is unmatchedwith any other SCX phases.0 2 4 6 8 10 12 14minColumn: Proteomix SAX-NP5 (5 µm, 4.6x50 mm)Mobile phase: A, 20 mM Tris, pH 8.0B, A + 0.5 M NaClGradient: 0-50%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 nmFigure 8 is a typical test chromatogram for a 5 µm,Proteomix SAX-NP column for separation of a mixture ofovalbumin and BSA. High resolution and high selectivity ofProteomix SAX-NP phase can well separate the impuritiescontained in ovalbumin sample, as well as the BSA dimerfrom BSA.www.sepax-tech.com 4 1-877-SEPAX-US
Page 3: Proteomix ® Ion-exchange PhasesGen
Page 7 and 8: High Lot-to-Lot ReproducibilityWith
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

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