Ion Exchange Chromatography COSMOGEL IEX Series Columns

Transcription

1 Ion Echange Chromatography COSMOGEL IEX Series Columns Available in 3 different ion-echange modes Anion-echange type, Cation-echange type, Amphoteric ion-echange type Available for 3 different application areas for Purification, for Ultra-fast analysis, for Precise analysis For separation of biopolymers such as proteins or nucleic acids COSMOGEL IEX Series COSMOGEL IEX Series are available in amphoteric ion-echange type in which two kinds of packing materials are mied, as well as in widely used anion-echange type and cation-echange type Ion Echange Mode Anion-echange Type Cation-echange Type Amphoteric ion-echange Type Packing Material Type Q Type Q-N Type S Type S-N Type M Type M-N Gel / Average Particle Size Hydrophilic Polymer / 5 µm Average Pore Size 000 Non-porous 000 Non-porous 000 Non-porous Functional Group -CH 3 N + (CH 3 ) 3 -(CH ) 3 SO 3 - -CH 3 N + (CH 3 ) 3 + -(CH ) 3 SO 3 - Protein Binding Capacity 0-50 mg -0 mg mg 0-8 mg BSA/ml-resin human IgG/ml-resin mg(bsa)/ml mg(igg)/ml 6-0 mg(bsa)/ml 5-9 mg(igg)/ml Target Sample Acidic Proteins / DNA Basic Proteins All Proteins Application For Purification For Ultra-fast For Precise For Purification For Ultra-fast For Precise For Purification For Precise Column Size ID Length(mm) Comprehensive isolation of proteins by Amphoteric ion-echange type (Type M) The amphoteric ion-echange type enables the simultaneous separation of both acidic and basic proteins in one application 3,4,

2 Separation by different ion-echange mode of amphoteric ion-echange type (Type M) Amphoteric ion-echange type can be used as anion-echange type or cation-echange type Application Types COSMOGEL IEX Series are available for 3 types of applications: purification, ultra-fast analysis and precise analysis For Purification: Type Q, Type S, Type M (Packing Material: Porous, Column Size: 46 mm ID 50 mm) Type Q, Type S and Type M with porous packing materials have higher binding capacity of proteins than the respective nonporous type, which means that peak shape does not spread even with injection of a large volume of sample Therefore they are highly suitable for purification of large sample 3 W / (sec) (Ribonuclease A) Type S Type S-N Sample loading amount (µg) For Precise : Type Q-N, Type S-N, Type M-N (Packing Material: Non-porous, Column Size: 46 mm ID 00 mm) Type Q-N, Type S-N, Type M-N with non-porous packing materials reduce spreading of samples in packing materials, result in high resolution separation for precise analysis such as quality control of antibody drugs The longer column length also contributes to the sharper peaks 3 3

3 For Ultra-fast : Type Q-N, Type S-N, Type M-N (Packing Material: Non-porous, Column Size: 46 mm ID 30 mm) The resolution with non-porous packing materials is not much affected by high flow rate and thus the materials are suitable for fast analysis The shorter column length contributes to the fast analysis Application Data Urease by Type M Ecoli Proteins Column: COSMOGEL IEX Type M Column size: 46mmID 50mm Mobile phase: A: 0mmol/l Tris buffer(ph83) B: 0mmol/l Tris buffer(ph83) + 05mol/l NaCl B conc 000% 30min Linear gradient Flow rate: 05 ml/min Temperature: 30 C Detection: UV80nm Sample: Urease (00g) Column: COSMOGEL IEX Type Q Column size: 46mmID 50mm Mobile phase: A: 0mmol/l Tris buffer(ph83) B: 0mmol/l Tris buffer(ph83) + 05mol/l NaCl B conc 000% 30min Linear gradient Flow rate: 05 ml/min Temperature: 30 C Sample: Ecoli Proteins Purification of Lysozyme from Egg White Ultra-fast of IgG

4 IgG by TypeQ-N IgG by TypeS-N DNA (00bp Ladder) Oligonucleotide Ordering Information Ion Echange Mode Product Name Application Column Size Product Number Anion-echange Type Cation-echange Type COSMOGEL IEX Type Q For Purification 46 mm ID 50 mm COSMOGEL IEX Type Q-N For Ultra-fast 46 mm ID 30 mm COSMOGEL IEX Type Q-N For Precise 46 mm ID 00 mm COSMOGEL IEX Type S For Purification 46 mm ID 50 mm COSMOGEL IEX Type S-N For Ultra-fast 46 mm ID 30 mm 065- COSMOGEL IEX Type S-N For Precise 46 mm ID 00 mm COSMOGEL IEX Type M For Purification 46 mm ID 50 mm Amphoteric ion-echangetypee COSMOGEL IEX Type M-N For Precise 46 mm ID 00 mm Column Material : PEEK Connection : Waters Type Nijo Karasuma, Nakagyo-ku, Kyoto JAPAN TEL : +8-(0) FAX : +8-(0) Website : wwwnacalaicom infointl@nacalaicom For research use only, not intended for diagnostic or drug use 0

5 I HPLC Columns II UHPLC Columns III Preparative Packing Materials IV Related Products CH3 (CH33 -(CH3SO3 - -CH33 3 -(CH3SO mg PEEK Anion- - Cation V Applications VI Technical Notes - Column: COSMOGEL IEX Column size: 46mmID-50mm Mobile phase: A: 0mmol/l Phosphate buffer(ph68) B: 0mmol/l Phosphate buffer(ph68) + 05mol/l NaCl B conc 000% 30min Linear gradient Flow rate: 05 ml/min Temperature: 30C Sample: ; Ribonuclease A (5g) ; Cytochrome C (5g) 3; Trypsin Inhibitor (60g) 4; -Galactosidase (60g) Type Q Type S Type M AP-74 VII Inde

6 - Column: COSMOGEL IEX Type S Column size: 46mmID-50mm Mobile phase: A: 0mmol/l Phosphate buffer(ph68) B: 0mmol/l Phosphate buffer(ph68) + 05mol/l NaCl B conc 000% 30min Linear gradient Flow rate: 05 ml/min Temperature: 30C Detection: UV80nm Sample: ; Ribonuclease A ; Cytochrome C 3; Lysozyme Each protein 500g 00g 0g - Column: COSMOGEL IEX Column size: Mobile phase: A: 0mmol/l Phosphate buffer(ph68) B: 0mmol/l Phosphate buffer(ph68) + 05mol/l NaCl B conc 000% 30min Linear gradient Flow rate: 05 ml/min Temperature: 30C Sample: ; Ribonuclease A (65g) ; Cytochrome C (65g) 3; Lysozyme (65g) Type S (46mmID-50mm) -- Column: COSMOGEL IEX Column size: Mobile phase: A: 0mmol/l Phosphate buffer(ph68) B: 0mmol/l Phosphate buffer(ph68) + 05mol/l NaCl B conc 000% Linear gradient Type S: 30min Type S-N: 0min Flow rate: Type S: 05 ml/min Type S-N: 0 ml/min Temperature: 30C Sample: ; Ribonuclease A (65g) ; Cytochrome C (65g) 3; Lysozyme (65g) Type S (46mmID-50mm) Type S-N (46mmID-00mm) Type S-N (46mmID-30mm) AP-85 AP-8 AP-83 I HPLC Columns II UHPLC Columns III Preparative IV Related Products V Applications VI Technical Notes VII Inde Packing Materials

7 I HPLC Columns II UHPLC Columns Column: Column size: Mobile phase: A: 0mmol/l Tris buffer(ph83) B: 0mmol/l Tris buffer(ph83) + 05mol/l NaCl B conc 0% (0-3min) 000% (3-43min) 00% (43-53min) Flow rate: 05 ml/min Temperature: 8C Equipment ÄKTAeplorer 0 S Sample: Trypsin Inhibitor from Soybean (mg/ml) InjVol 500l Data courtesy of Kei WADA, Ph D, Department of Biological Sciences, Graduate School of Science, Osaka University mau COSMOGEL IEX Type Q (46mmID-50mm) Competitor (5mmID-50mm) Time(min) AP-95 III Preparative Packing Materials IV Related Products V Applications VI Technical Notes Ordering Information Anion- Cation VII Inde mau Time(min) Column: COSMOGEL IEX Type Q Column size: 46mmID-50mm Mobile phase: A: 0mmol/l Tris buffer(ph80) B: 0mmol/l Tris buffer(ph80) + 05mol/l NaCl B conc 30% (0min) 35% (0-30min) 50% (30-50min) 00% (50-55min) 30% (55min-) Flow rate: 05 ml/min Temperature: room temperature Detection: UV80nm Sample: InjVol His-tag Protein in 0mmol/l Tris buffer(ph80) (Lane 4) 50ml Lane : Protein Makers for SDS-PAGE (# ) Lane : His-tag protein [pet-4b(+)] in E Coli lysate Lane 3: Purified fraction by COSMOGEL His-Accept SF Lane 4: Desalted fraction Lane 5: Purified fraction by COSMOGEL IEX Type Q AP-97

8 COSMOGEL IEX Type Q Comparison Data COSMOGEL IEX Series are available for 3 different applications, for purification, for ultra-fast analysis and for precise analysis COSMOGEL IEX Type Q is highly suitable for purification of large samples The followings are comparison data of COSMOGEL IEX Type Q and competitors columns using a widely used equipment, ÄKTA eplorer 0S of GE Healthcare Comparison of Separation Performance COSMOGEL IEX Type Q shows higher resolution compared to competitors columns under the same condition Furthermore COSMOGEL IEX Type Q enables high purity purification for impurity samples COSMOGEL IEX Type Q (46mmID-50mm) Competitor A- (5mmID-50mm) Mobile phase: A: 0mmol/l Tris buffer(ph83) B: 0mmol/l Tris buffer(ph83) + 05mol/l NaCl B conc 0% (0-3min) 000% (3-43min) 00% (43-53min) Flow rate: 05 ml/min Temperature: 8 C Equipment: ÄKTAeplorer 0 S Sample: Trypsin Inhibitor from Soybean (mg/ml) InjVol : 500 µl COSMOGEL IEX Type Competitor A- High Resolution! Data courtesy of Kei WADA, Ph D, Department of Biological Sciences, Graduate School of Science, Osaka University

9 Comparison of Pressure The following table shows pressure comparison of COSMOGEL IEX Type Q and competitors columns under the same condition using equipment, ÄKTA eplorer 0S of GE Healthcare Although the pressure of COSMOGEL IEX Series is a little higher, COSMOGEL IEX Type Q can be an alternative competitors columns Column Name Column Size Pressure underthe same condition COSMOGEL IEX Type Q 46 mm ID 50 mm -5 MPa COSMOGEL IEX Type Q-N 46 mm ID 00 mm MPa Competitor A- 50 mm ID 50 mm 6-8 MPa Competitor A- ml MPa Mobile Phase: A: 0 mmol/l Tris buffer(ph 83) B: 0 mmol/l Tris buffer(ph 83) + 05 mol/l NaCl B conc 0% (0-3 min) 0 00% (3-43 min) 00% (43-53 min) Flow Rate: 05 ml/min Temperature: 8 C Detection: UV0 nm Equipment: ÄKTA eplorer 0 S Sample: Trypsin Inhibitor from Soybean ( mg/ml) Injection Vol: 500 l Data courtesy of Kei WADA, Ph D, Department of Biological Sciences, Graduate School of Science, Osaka University Accessories such as sample loop may not suitable if their pressure resistance is lower than the equipment used Please refer to applicable instructions Characteristic of Each Column Column Name Column Size Average Particle Size Average Pore Size Maimum Flow Rate Pressure Limit COSMOGEL IEX Type Q 46 mm ID 50 mm 5 m ml/min or less appro MPa * COSMOGEL IEX Type Q-N 46 mm ID 00 mm 5 m Non-Porous 0 ml/min or less appro 0 MPa * Competitor A- 50 mm ID 50 mm 0 m N/A 30 ml/min or less 4 MPa Competitor A- ml 5 m N/A 00 ml/min or less 5 MPa * Maimum Flow Rate of COSMOGEL IEX Series is specified in the Instruction When flow rate is 08 ml/min, pressure is appro MPa * Maimum Flow Rate of COSMOGEL IEX Series is specified in the Instruction When flow rate is 0 ml/min, pressure is appro 0 MPa ÄKTA eplorer is registered trademarks ofge Healthcare Nijo Karasuma, Nakagyo-ku, Kyoto JAPAN TEL : +8-(0) FAX : +8-(0) Website : wwwnacalaicom infointl@nacalaicom For research use only, not intended for diagnostic or drug use 07