Sepax SRT -C, Zenix -C SEC Phases

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1 Sepax SRT -C, Zenix -C SEC Phases Complimentary Phases to SRT for Derivatized Monoclonal Antibodies General Description Stationary Phase Structure SRT, Zenix, SRT-C and Zenix-C SEC phases developed based on innovative surface coating technology comprised of uniform, hydrophilic, and neutral nanometer thick films chemically bonded on high purity and mechanically stabilized silica. The two different types of coating chemistries, SRT and Zenix, stand-up monolayer bonded on porous silica, and SRT-C and Zenix-C, lay-down monolayer on porous silica offer ideal phase chemistries for sample type specific separation. The um based Zenix and Zenix- C, and um based SRT and SRT-C allow for high resolution and performance separation. The combination of these four lines of SEC phases provides a powerful total solution for robust, reproducible and highest resolution, size based separations of biological molecules. Figure. Phase structure difference: a monolayer stands up on the silica surface for SRT and Zenix, and a monolayer lays down on the silica surface for SRT-C and Zenix-C. Difference in Particle Size Featured Characteristics Higher resolution over wider MW range High lot-to-lot reproducibility High protein recovery with intact biological activity Negligible non-specific interactions Ideal for hydrophobic proteins, and monoclonal antibodies derivatized with polymer branches Suitable for separation and analysis of general biological samples Zenix & Zenix-C µm SRT & SRT-C µm Key features of Sepax SEC phases Figure. Zenix and Zenix-C are based on µm porous silica. SRT and SRT-C are based on µm porous silica. Characteristics SRT Zenix SRT-C Zenix-C Particle size µm µm µm µm Pore size (Å) Resolution 00, 0, 00, 00, 000 & 000 High 00, 0, 00 0, 00 & 00 00, 0, 00 Highest, Short column for faster separation High Highest, Short column for faster separation Efficiency* High Doubled from um High Doubled from um Selectivity Same for SRT and Zenix Same for SRT-C and Zenix-C Surface structure Chemically bonded stand-up monolayer Chemically bonded lay-down monolayer Recommended Sample Types Monoclonal antibodies, proteins, peptides, nucleic acids, oligonucleotides, virus, and water-soluble polymers Tough samples such as hydrophobic proteins like insulin, membrane proteins monoclonal antibodies derivatized with polymer branches, e.g. polypeptide, PEG. * Using Ribonuclease A as the test molecule on a 7.8x00mm column, the plate count is,000 for Zenix vs.,0 for SRT

2 Protein MW Calibration Protein molecular weight vs elution volume is plotted in Figure and Figure, indicating that SRT C-0, 00 and 00, and Zenix-C 00, 0 and 00 have large linear elution regions for better resolution over a wider MW range. Figure. Protein MW calibration with elution volume for SRT-C phases. Applications Figure. Separation of a protein mixture by SRT-C-00 column. 7 LogMW SRT C-0 SRT C-00 SRT C Elution Volume (ml) Columns: 7.8x00 mm, µm Mobile phase: 0 mm Sodium Phosphate, ph 7.0 Flow rate:.0 ml/min Detection: UV nm Injection volume: 0 µl Sample:. Thyroglobulin, 70 kd;. -Globulin, 8 kd;. BSA, kd;. Ovalbumin, kd;. Myoglobin, 7. kd;. Ribonuclease A,.7 kd; 7. B,. kd; 8. Uracil, 0 Figure. Protein MW calibration with elution volume for Zenix-C phases. LogMW 7 Zenix C-00 Zenix C-0 Zenix C Elution Volume (ml) Columns: 7.8x00 mm, µm Mobile phase: 0 mm phosphate buffer, ph 7.0 Flow rate:.0 ml/min Detection: UV nm Injection volume: 0 µl Sample:. Thyroglobulin, 70 kd;. -Globulin, 8 kd;. BSA, kd;. Ovalbumin, kd;. Myoglobin, 7. kd;. Ribonuclease A,.7 kd; 7. B,. kd; 8. Uracil, Column: SRT-C-00 (7.8x00 mm, µm) Mobile phase: 0 mm Sodium Phosphate, ph 7 Flow rate:.0 ml/min Temperature: ambient (~ C) Detection: UV nm Injection: 0 µl Sample: ) Thyroglobulin, 70kD; ) -Globulin, 8 kd; ) Ovalbumin, kd; ) Myoglobin, 7. kd; ) Poly-DLalanine (- kd); ) Uracil, 0D. Figure. Separation of monoclonal antibody and its high MW species by SRT-C-00 column A Column: SRT-C-00 (7.8x00 mm, µm) Mobile phase: 0 mm Sodium Phosphate, ph 7 Flow rate:.0 ml/min Temperature: Ambient (~ C) Detection: UV nm Injection: 0 µl Sample: Monoclonal antibody (. mg/ml) Figure 7. Separation of a protein mixture by Zenix-C 00 column Column: Zenix-C 00 (7.8x00 mm, µm) Mobile phase: 0 mm Sodium phosphate, ph

3 Flow rate:.0 ml/min Temperature: ambient (~ C) Detection: UV nm Injection: 0 µl Sample: ) Thyroglobulin, 70kD; ) -Globulin, 8 kd; ) Ovalbumin, kd; ) Myoglobin, 7. kd; ) Poly-DLalanine (- kd); ) B,.KD. Figure 8. BSA loading test on a Zenix-C SEC 00 column Column: Zenix SEC-00 ( µm, 7.8x00 mm) Mobile phase: 0 mm Sodium phosphate, ph 7.0 Flow Rate:.0 ml/min Injection volume: 0 µl BSA concentration:,, 0, and 0 mg/ml (from low to high) Detection: UV nm Figure 9. Separation of Biorad protein mix on Zenix-C SEC 00, 0 and 00 columns. Zenix C-00 Zenix C-0 Zenix C-00, Columns: Zenix-C ( µm, 7.8x00 mm) Mobile phase: 0 mm Sodium phosphate, ph 7.0 Flow rate:.0 ml/min Detection: UV nm Injection: 0 µl Sample: ) Thyroglobulin, 70 kd; ) -Globulin, 8 kd; ) Ovalbumin, kd; ) Myoglobulin,.9 kd; ) Vitamin B, D. Figure 0. Separation of protein mixture A by Zenix-C SEC 00, 0 and 00 columns. Zenix C-00 Zenix C-0 Zenix C-00, Column: Zenix-C ( µm, 7.8x00 mm) Mobile phase: 0 mm Sodium phosphate, ph 7.0 Flow rate:.0 ml/min Temperature: Ambient (~ C) Detection: UV nm Injection: 0 µl ( L for.x00 mm) Sample: ) Thyroglobulin (.0 mg/ml), 70 kd; ) BSA (.0 mg/ml), kd; ) Ribonuclease A (.0 mg/ml),.7 kd, and ) Uracil (. g/ml), 0D. Figure. Elution of insulin from Zenix-C 00 column Columns: Zenix SEC ( µm, 7.8x00 mm) Mobile phase: 0 mm Sodium phosphate, ph 7.0 Flow rate:.0 ml/min Detection: UV nm Injection: 0 µl Sample: Insulin (from a commercial source containing impurity)

4 Dimer BSA (N=,0) SRT SEC-00 BSA (N=,090) Unique benefits of Zenix and Zenix-C Phases Zenix and Zenix-C columns offer highest efficiency and resolution for biomolecules over a wide MW range. Aggregate Dimer Zenix SEC Figure. Column: 7.8x00mm; 0 mm sodium phosphate, ph 7.0; Flow rate:.0 ml/min; UV nm; Injection: 0 L (.0 mg/ml). SRT SEC Technical Specifications Phase SRT-C-0 SRT-C-00 SRT-C-00 Material Particle size m m m Pore size (Å) ~ 0 ~ 00 ~ 00 Protein MW range (native) 00-0,000,000,0,000,000,000,000 ph stability 8. (ph can be 8. (ph can be Backpressure ( ml/min, 7.8x00mm) ~ 700 ~ 700 ~ 700 Maximum backpressure (psi) ~,00 ~,00 ~, (ph can be Salt concentration range 0 mm -.0 M 0 mm -.0 M 0 mm -.0 M Maximum temperature ( o C) ~ 80 ~ 80 ~ 80 Mobile phase compatibility Aqueous and organic Aqueous and organic Aqueous and organic Zenix SEC Technical Specifications Phase Zenix SEC-00 Zenix SEC-0 Zenix SEC-00 Material Particle size µm µm µm Pore size (Å) ~ 00 ~ 0 ~ 00 Protein MW range (native) 00-00, ,000,000,0,000 ph stability Backpressure for 7.8x00 mm (.0 ml/min) Backpressure for.x00 mm (0. ml/min) 8. (ph can be 8. (ph can be ~,00 psi ~,7 psi ~,00 psi ~,00 psi ~,0 psi ~,000 psi Maximum backpressure (psi) ~,00 ~,00 ~,00 8. (ph can be Salt concentration range 0 mm -.0 M 0 mm -.0 M 0 mm -.0 M Maximum temperature (oc) ~ 80 ~ 80 ~ 80 Mobile phase compatibility Aqueous and organic Aqueous and organic Aqueous and organic

5 Ordering Information: Distributed by: The Nest Group, Inc.. Call for pricing and availability. Other dimension and pore size available upon request SRT-C-0 ( m, 0Å) 7.8x00 T x0 (Guard) T x00 T 0-0.x0 (Guard) T 0-0 0x00 T x00 T 0-0 SRT-C-00 ( m, 00Å) Zenix Zenix-C T Z Includes Zenix (00Å, 7.8x00mm) Zenix-C (00Å, 7.8x00mm) Zenix & Zenix-C SEC Solution Pack T Z0-780 Includes Zenix (0Å, 7.8x00mm) Zenix-C (0Å, 7.8x00mm) 7.8x00 T x0(Guard) T x00 T 00-0.x0 (Guard) T x00 T x00 T 00-0 Limited Time Introductory Offer $,00 / pk. SRT-C-00 ( m, 00Å) 7.8x00 T x0(Guard) T x00 T 00-0.x0 (Guard) T x00 T x00 T 00-0 Zenix-C-0 ( m, 0Å) 7.8x00 T x0(Guard) T x00 T 0-0.x0 (Guard) T 0-0 0x00 T x00 T 0-0 Zenix-C-00 ( m, 00Å) 7.8x00 T x0 (Guard) T x00 T 00-0.x0 (Guard) T x00 T x00 T 00-0 Zenix-C-00 ( m, 00Å) 7.8x00 T x0(Guard) T x00 T 00-0.x0 (Guard) T x00 T x00 T

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