Tosoh Bioscience llc Reversed Phase Chromatography TSK-GEL Reversed Phase Columns Tosoh Bioscience offers a broad line of analytical and preparative TSK-GEL columns for the Reversed Phase Chromatography (RPC) analysis of small molecular weight drugs, amino acids, carbohydrates, lipids, nucleic acid fragments, peptides and proteins. For high-speed separations we recommend the porous, silica-based TSK-GEL Super Series and the nonporous, polymeric NPR columns. For greater stability at high ph or to benefit from alternative selectivities, we recommend either the silica-based 80T S columns or polymer-based TSK-GEL columns. TSK-GEL 80T S columns are stable up to ph 7.5. The best peak shapes for basic compounds and extra retention and selectivity for polar compounds are obtained with the recently introduced TSKgel ODS-00V and TSKgel ODS-00Z Reversed Phase Columns. Properties of Silica-based TSK-GEL RPC Columns TSKgel Bonded Functional End Particle Carbon Pore Exclusion Application/ Features Column Phase Group Capping Size (µm) Load % Size (Å) Limit (Da)* ODS-80TM Monomeric C8 High 5, 0, 0 5 80 6,000 Low MW peptides, peptide mapping, pharmaceuticals, nucleic acid fragments ODS-80TS Monomeric C8 Complete 5, 0, 0 5 80 6,000 As ODS-80TM, but more stable at higher ph Octyl-80TS Monomeric C8 Complete 5 80 6,000 As ODS-80TM, but faster kinetics than ODS. Lower hydrophobic selectivity. CN-80TS Monomeric Cyano Complete 5 8 80 6,000 Alternative to ODS and Octyl columns for analysis of polar compounds ODS-0T Polymeric C8 High 5, 0, 0 0 0,000 Expands ODS-80TM applications for peptides and small proteins ODS-0A Polymeric C8 None 5, 0 0 0 0,000 Polyaromatic hydrocarbons Super-ODS Polymeric C8 Complete 8 0 0,000 High-throughput analyses of peptides, tryptic digests and low MW Super-Octyl Polymeric C8 Complete 5 0 0,000 pharmaceuticals, purines and pyrimidines, nucleosides, nucleotides Super-Phenyl Polymeric C 6H 5 Complete 0 0,000 OligoDNA RP Monomeric C8 None 5 0 50 65,000 Specialty column for analysis and preparative purification of oligonucleotides, RNA and DNA-fragments TMS-50 Monomeric C Complete 0 5 50 00,000 Specialty columns for protein separations ODS-00V Monolayer C8 Complete 5 5 00 Stronger retention for polar compounds ODS-00Z Monolayer C8 Complete 5 0 00 Maximum hydrophobic retention Properties of Polymer-based TSK-GEL RPC Columns TSKgel Bonded Functional Particle Pore Exclusion Application/ Features Column Phase Group Size (µm) Size (Å) Limit (Da) * Octadecyl-PW Monomeric C8 5 5 8,000 Low MW peptides and pharmaceuticals unstable at low ph Octadecyl-PW Monomeric C8 7 500 00,000 Medium and high MW peptides and proteins especially if unstable at low ph Phenyl-5PW RP Monomeric C 6 H 5 0 000,000,000 High MW peptides and proteins. Phenyl group modifies selectivity. Octadecyl-NPR Monomeric C8.5 Nonporous >,000,000 Rapid separation of high MW peptides and proteins * for globular proteins 50 Phenomenex
TSK-GEL Reversed Phase Columns (cont'd) Protein Column: TSKgel Octadecyl-NPR Dimensions:.6 mm x.5 cm Part No.: CH0-7967 Mobile Phase: 0 min linear gradient from 5 % to 80 % Acetonitrile in 5 mm HCIO Flow Rate:.5 ml/min Detection: UV @ 0 nm Sample: A. Blank injection, B. 50 ng each of:. Ribonuclease A. Lysozyme. Insulin 5. Transferrin. Cytochrome c 6. Myoglobin Peptides Column: Dimensions: Part No.: Mobile Phase: Flow Rate: Detection: Temperature: Sample: TSKgel Octadecyl-PW.6 mm x 5 cm CH0-7968 0 min linear gradient from 0. % TFA/Acetonitrile from 90/0 to 0/70.0 ml/min UV @ 5 nm Ambient 50 ng each of:. Met-Enkephalin 5. Bombesin. Bradykinin 6. Angiotensin I. Leu-Enkephalin 7. Somatostatin. Neurotensin 8. Insulin (Bovine) B A 5 6 6 7 8 5 0 6 min 0 0 0 min Comparison of Three Reversed Phase Columns Columns: A: TSKgel ODS-80T S B: TSKgel Octyl-80T S A C: TSKgel CN-80T S Dimensions:.6 mm x 5 cm Mobile Phase: 70 % Methanol (except CN-80T S with 50 % Methanol) Flow Rate:.0 ml/min Temperature: 5 C Sample: 50 ng each of:. Uracil. Toluene. Benzene. Naphthalene 0 6 8 min B C 0 6 8 min 0 6 8 min Phenomenex 5
Tosoh Bioscience llc TSK-GEL Reversed Phase Columns (cont'd) 80T S Series Stainless Steel Columns I I I I CH0-79 850 ODS-80T S, 80 Å.0 5.0 5 CH0-79 85 ODS-80T S, 80 Å.0 5.0 5 CH0-79 700 ODS-80T S, 80 Å.6 7.5 5 CH0-795 70 ODS-80T S, 80 Å.6 5.0 5 CH0-796 70 ODS-80T S, 80 Å.6 5.0 5 CH0-797 665 ODS-80T M, 80 Å.6 7.5 5 CH0-798 088 ODS-80T M, 80 Å.6 5.0 5 CH0-799 089 ODS-80T M, 80 Å.6 5.0 5 CH0-790 00 ODS-80T M, 80 Å 7.8 0.0 0 CH0-79 7 Octyl-80T S, 80 Å.6 5.0 5 CH0-79 75 Octyl-80T S, 80 Å.6 5.0 5 CH0-79 78 CN-80T S, 80 Å.6 5.0 5 CH0-79 79 CN-80T S, 80 Å.6 5.0 5 CH0-8 * 95 ODS-80T S Guard cartridge, /pk.0.0 5 AF0-798 00 Pre-column filter CH0-8 * 908 Guard cartridge holder.0 ODS-00 Series Stainless Steel Columns CH0-87 55 ODS-00V, 00 Å.6 5.0 5 CH0-88 56 ODS-00V, 00 Å.6 5.0 5 CH0-89 6 ODS-00Z, 00 Å.6 5.0 5 CH0-850 6 ODS-00Z, 00 Å.6 5.0 5 CH0-85 5 ODS-00V Guard cartridge, /pk..5 5 CH0-85 5 ODS-00Z Guard cartridges, /pk..5 5 CH0-85 908 Guard cartridge holder ODS-0 Series Stainless Steel Columns CH0-799 0766 ODS-0A, 0 Å.6 5.0 5 CH0-790 07 ODS-0A, 0 Å.6 5.0 5 CH0-79 079 ODS-0A, 0 Å 7.8 0.0 0 CH0-79 067 ODS-0A, 0 Å.5 0.0 0 CH0-79 85 ODS-0T, 0 Å.0 5.0 5 CH0-79 85 ODS-0T, 0 Å.0 5.0 5 CH0-795 0767 ODS-0T, 0 Å.6 5.0 5 CH0-796 075 ODS-0T, 0 Å.6 5.0 5 CH0-797 070 ODS-0T, 0 Å 7.8 0.0 0 CH0-798 07 ODS-0T, 0 Å.5 0.0 0 CH0-799 07 ODS-0T, 0 Å 55.0 0.0 5-5 CH0-7950 07 ODS-0T, 0 Å 55.0 60.0 5-5 CH0-8 * 95 ODS-0T Guard cartridge, /pk.0.0 5 CH0-795 076 ODS-0T/A Guard column.5 7.5 0 CH0-8 * 908 Guard cartridge holder.0.0 Super Series Stainless Steel Columns I I I I CH0-795 95 Super-ODS, 0 Å.0 5.0 CH0-795 95 Super-ODS, 0 Å.0 0.0 CH0-7955 85 Super-ODS, 0 Å.6 5.0 CH0-7956 897 Super-ODS, 0 Å.6 0.0 CH0-7957 875 Super-Octyl, 0 Å.6 5.0 CH0-7958 876 Super-Octyl, 0 Å.6 0.0 CH0-7959 877 Super-Phenyl, 0 Å.6 5.0 CH0-7960 878 Super-Phenyl, 0 Å.6 0.0 CH0-796 967 Guard cartridge /pk.0.0 CH0-8 * 908 Guard cartridge holder.0 CH0-796 ** 807 Guard cartridge /pk.0 0. CH0-796 806 Cartridge holder (for Part No. CH0-796) Oligo DNA and TMS Series Stainless Steel Columns CH0-796 5 OligoDNA RP, 50 Å.6 5.0 5 CH0-7965 5 OligoDNA RP, 50 Å 7.8 5.0 5 CH0-7966 0790 TMS-50, 50 Å.6 7.5 0 NPR and PW-RP Series Stainless Steel Columns CH0-7967 005 Octadecyl-NPR nonporous.6.5.5 CH0-7968 7500 Octadecyl-PW, (00-00 Å).6 5.0 5.0 CH0-7969 750 Octadecyl-PW, (00-00 Å) 6.0 5.0 5.0 CH0-7970 875 Octadecyl-PW, (00-00 Å).0 5.0 5.0 CH0-797 5 Octadecyl-PW, 500 Å.6 5.0 7.0 CH0-797 657 Octadecyl-PW, 500 Å.5 5.0.0 CH0-797 8755 Octadecyl-PW, 500 Å.0 5.0 7.0 CH0-797 658 Octadecyl-PW, 500 Å 55.0 0.0 0.0 CH0-7975 8756 Phenyl-5PW RP, 000 Å.0 7.5 0.0 CH0-7976 080 Phenyl-5PW RP, 000 Å.6 7.5 0.0 CH0-7977 660 Phenyl-5PW RP, 000 Å.5 5.0.0 CH0-7978 66 Phenyl-5PW RP, 000 Å 55.0 0.0 0.0 CH0-7979 59 Phenyl-5PW RP Cartridge, /pk.0.0 CH0-798 66 Phenyl-5PW RP Guard column 5.0 5.0 CH0-798 6 Octadecyl-PW Cartridge.0.0 CH0-798 750 Octadecyl-PW Guard column.6.0 CH0-7985 750 Octadecyl-PW Guard column 6.0.0 CH0-7986 60 Octadecyl-PW Cartridge, /pk.0.0 CH0-7987 t 679 Octadecyl-PW Prep Guardgel Kit CH0-7988 659 Octadecyl-PW Guard Column 5.0 5.0 Guard Columns CH0-8 * 908 Guard cartridge holder.0 * For.0 mm ID cartridges ** For.6 mm ID columns t For.5 mm ID columns For 55 mm ID columns 5 Phenomenex
Normal Phase Chromatography TSK-GEL Normal Phase Columns Separate hydrophilic compounds such as peptides, carbohydrates, and small polar molecules for drug discovery. The TSKgel Amide-80 column offers an excellent alternative to amino bonded stationary phases and consists of 5 or 0 µm silica particles with 80 Å pores in a stainless steel format. Polyalcohols Column: TSKgel Amide-80 Dimensions:.6 mm x 5 cm Part No.: CH0-7995 Mobile Phase: Acetonitrile/Water (75:5) Flow Rate:.0 ml/min Temperature: 5 C Detection: RI Sample: A. Ethylene Glycol 0 µm, 0 µl D. Xylitol 0 µm, 0 µl B. Glycerine 0 µm, 0 µl E. Mannitol 0 µm, 0 µl C. Erythritol 0 µm, 0 µl F. Inositol µm, 0 µl B C D E Normal Phase Amide-80 Series Stainless Steel Columns I I I I CH0-7989 969 Amide-80.0 5.0 5 CH0-7990 9695 Amide-80.0 0.0 5 CH0-799 9696 Amide-80.0 5.0 5 CH0-799 9697 Amide-80.0 5.0 5 CH0-799 95 Amide-80.6 5.0 5 CH0-799 95 Amide-80.6 0.0 5 CH0-7995 07 Amide-80.6 5.0 5 CH0-7996 59 Amide-80 7.8 0.0 0 CH0-7997 60 Amide-80.5 0.0 0 Guard Columns CH0-7998 90 Amide-80 Guard column.6.0 5 CH0-7999 6 Amide-80 Guard column.5 7.5 0 CH0-8000 0 Amide-80 Guard cartridge, /pk.0.0 5 CH0-8 908 Guard cartridge holder (For all mm ID columns) A F 0 0 0 min Phenomenex 5
Size Exclusion Chromatography - SW-Type TSK-GEL SW-type SW, SW XL, SuperSW Silica-based for aqueous GFC of proteins and peptides Properties and Molecular Weight Separation Ranges for TSK-GEL SW-type Packings Molecular weight of sample (Da) TSKgel Particle Pore Globular Dextrans Polyethylene Column Size (µm) Size (Å) proteins glycols and oxides SuperSW000 5 5,000.5 x 0 5,000 x 0 500 5,000 G000SWXL/BioAssist GSWXL 5 5 5,000.5 x 0 5,000 x 0 500 5,000 QC-PAK TSK 00 5 5 5,000.5 x 0 5,000 x 0 500 5,000 G000SW 0, 5 5,000 x 0 5,000 x 0 500 5,000 SuperSW000 50 x 0 5 x 0 5,000 7 x 0,000.5 x 0 G000SWXL/BioAssist GSWXL 5 50 x 0 5 x 0 5,000 7 x 0,000.5 x 0 QC-PAK TSK 00 5 50 x 0 5 x 0 5,000 7 x 0,000.5 x 0 G000SW 0 50 50 x 0 5 x 0 5,000 7 x 0,000.5 x 0 G000SWXL/BioAssist GSWXL 8 50 x 0 7 x 0 6,000 5 x 0 5,000.5 x 0 5 G000SW, 7 50 x 0 7 x 0 6,000 5 x 0 5,000.5 x 0 5 Data generated using the following conditions: Columns: Two µm,.6 mm x 0 cm TSK-GEL SuperSW columns in series; two 5 µm, 7.8 mm x 0 cm TSK-GEL SW XL columns in series; two 0 µm, 7.5 mm x 60 cm TSK-GEL SW columns in series Mobile Phases: Globular proteins: 0. M NaCl in 0. M (0.05 M for SW XL columns) phosphate buffer, ph 7 Dextrans and polyethylene glycols and oxides (PEOs): distilled water Comparison of TSKgel SuperSW000 and TSKgel G000SW XL for the Separation of Proteins Columns: A: TSKgel G000SW XL B: TSKgel SuperSW000 Dimensions: A: 7.8 mm x 0 cm B:.6 mm x 0 cm Mobile Phase: 0. M Na SO in 0. M Phosphate Buffer with 0.05 % NaN, ph 6.7 Flow Rate: A:.0 ml/min B: 0.5 ml/min Temperature: 5 C Detection: Sample: UV @ 0 nm. Thyroglobulin, 0.5 mg/ ml (660,000 Da). Ribonuclease A,.5 mg/ ml (,600 Da). g-globulin,.0 mg/ ml (50,000 Da) 5. p-aminobenzoic Acid, 0.0 mg/ ml (7 Da). Ovalbumin,.0 mg/ ml (,000 Da) 50 0 A 50 0 B 5 mv 0 0 0 5 0 0 0 0 5 7 9 5 min 0 5 7 9 5 min 5 Phenomenex
Size Exclusion Chromatography - PW-Type TSK-GEL PW-type PW, PW XL Polymer-based for aqueous GFC of water soluble polymers Properties and Molecular Weight Separation Ranges for TSK-GEL PW-type Packings Molecular weight of sample (Da) TSKgel Particle Average Polyethylene Dextrans** Globular Column Size* (µm) Pore Size (Å) glycols and oxides proteins** G000PW 0 <00 up to,000 <,000 G000PW 0, 7, 0 5 up to,000 <5,000 G500PWXL 6 <00 up to,000 <8,000 G500PW 0, 7, 0 G000PWXL 6 00 up to 5 x 0 up to 6 x 0 500 8 x 0 5 G000PW 0, 7, 0 G000PWXL 0 500,000 x 0,000 7 x 0 5 x 0.5 x 0 6 G000PW 7, G5000PWXL 0 000,000 x 0 6 5 x 0.5 x 0 6 < x 0 7 G5000PW 7, 0, G6000PWXL/BioAssist G6PWXL >000 x 0 8 x 0 6 5 x 0 5 5 x 0 7 < x0 8 G6000PW 7, 5 GMPWXL <00 000 500 8 x 0 6 <5 x 0 7 < x 0 8 GMPW 7 G-Oligo-PW 6 5 up to,000 <,000 G-DNA-PW 0 >000 x 0 8 x 0 6 <5 x 0 7 < x 0 8 Data generated using the following conditions: Columns: TSK-GEL PW columns, 7.5 mm x 60 cm; TSKgel PW XL, G-Oligo-PW & G-DNA-PW, 7.8 mm x 0 cm Mobile Phases: Polyethylene glycols and oxides: distilled water; dextrans and proteins: 0. M phosphate buffer, ph 6.8 Flow Rates:.0 ml/min Note: * Larger particle sizes of each group are for.5 mm x 60 cm semi-preparative and 55 mm or 08 mm x 60 cm preparative columns. ** Maximum separation range determined from estimated exclusion limits. Polyethylene Glycol and Oxide Calibration Curves on TSK-GEL PW and TSK-GEL PW XL Columns Molecular Weight (Da) 0 6 0 5 0 0 0 A TSK-GEL PW Columns A G C B D E 0 5 0 F 0 6 0 5 0 0 0 B TSK-GEL PWXL Columns N M K J L H 5 0 5 TSKgel PW Columns Columns: A: TSK-GEL PW A: G000PW B: G500PW C: G000PW D: G000PW E: G5000PW F: G6000PW G: GMPW Dimensions: All 7.5 mm x 60 cm B: TSK-GEL PWXL H: G500PWXL J: G000PWXL K: G000PWXL L: G5000PWXL M: G6000PWXL N: GMPWXL Dimensions: All 7.8 mm x 0 cm Mobile Phase: Distilled Water Flow Rate:.0 ml/min Detection: RI Elution volume (ml) Elution volume (ml) Phenomenex 55
TSK-GEL SW- and PW-Type (cont'd) SW-Type Part No. Mfr. No. Description ID Length Particle Min. Number Flow Rate (ml/min) Maximum Price (mm) (cm) Size (µm) Theoretical Range Max. Pressure Plates Drop (kg/ cm ) Stainless Steel Columns I I I I I I I I CH0-800 867 SuperSW000.6 0 0,000 0. 0.5 0. 0 CH0-79 8675 SuperSW000.6 0 0,000 0. 0.5 0. 0 CH0-5 0850 G000SWXL 7.8 0 5 0,000 0.5.0. 70 CH0-5 085 G000SWXL 7.8 0 5 0,000 0.5.0. 70 CH0-5 085 G000SWXL 7.8 0 8 6,000 0.5.0. 5 CH0-800 65 QC-PAK GFC 00 7.8 5 5 0,000 0.5.0. 0 CH0-800 609 QC-PAK GFC 00 7.8 5 5 0,000 0.5.0. 0 CH0-0708 05788 G000SW 7.5 0 0 0,000 0.5.0. 0 CH0-07 05789 G000SW 7.5 0 0 0,000 0.5.0. 5 CH0-076 05790 G000SW 7.5 0 8,000 0.5.0. 5 CH0-800 050 G000SW 7.5 60 0 0,000 0.5.0. 0 CH0-07 050 G000SW 7.5 60 0 0,000 0.5.0. 50 CH0-8005 050 G000SW 7.5 60 6,000 0.5.0. 0 PEEK Columns CH0-86 007 BioAssist GSWXL 7.8 0 5 0,000 0.5.0. 70 CH0-8006 006 BioAssist GSWXL 7.8 0 5 0,000 0.5.0. 70 PW-Type Stainless Steel Columns CH0-8007 080 G-Oligo-PW 7.8 0 6,000 0.5 0.8.0 0 CH0-8008 080 G-DNA-PW 7.8 0 0 0,000 0. 0.5 0.6 0 CH0-078 0800 G500PW XL 7.8 0 6,000 0.5 0.8.0 0 CH0-075 080 G000PW XL 7.8 0 6,000 0.5 0.8.0 0 CH0-0756 080 G000PW XL 7.8 0 0 0,000 0. 0.8.0 0 CH0-0760 080 G5000PW XL 7.8 0 0 0,000 0. 0.8.0 0 CH0-076 080 G6000PW XL 7.8 0 7,000 0. 0.8.0 0 CH0-0767 0805 GMPW XL 7.8 0 7,000 0. 0.8.0 0 CH0-079 05760 G000PW 7.5 0 0 5,000 0.5.0. 0 CH0-07 0576 G000PW 7.5 0 0 5,000 0.5.0. 0 CH0-075 0808 G500PW 7.5 0 0 5,000 0.5.0. 0 CH0-079 0576 G000PW 7.5 0 0 5,000 0.5.0. 0 CH0-075 0576 G000PW 7.5 0 7,000 0.5.0. 0 CH0-0757 0576 G5000PW 7.5 0 7,000 0.5.0. 0 CH0-076 05765 G6000PW 7.5 0 7,000 0.5.0. 0 CH0-8009 0806 GMPW 7.5 0 7,000 0.5.0. 0 CH0-800 0505 G000PW 7.5 60 0 0,000 0.5.0. 0 CH0-80 0809 G500PW 7.5 60 0 0,000 0.5.0. 0 CH0-0750 0506 G000PW 7.5 60 0 0,000 0.5.0. 0 CH0-80 0507 G000PW 7.5 60 7 6,000 0.5.0. 0 CH0-80 0508 G5000PW 7.5 60 7 6,000 0.5.0. 0 CH0-80 0509 G6000PW 7.5 60 7 6,000 0.5.0. 0 CH0-805 0807 GMPW 7.5 60 7 6,000 0.5.0. 0 PEEK Columns CH0-806 00 BioAssist G6PW 7.8 0 7,000 0.5.0. 0 56 Phenomenex
Ion Exchange Chromatography Features and Benefits of Ion Exchange Columns BioAssist Fewer runs to collect required sample amounts Sharper peaks improve analysis and isolation Peak column housing reduces sample loss due to adsorption Easy scale-up Polymethacrylate particles Polymer-Based Ion Exchange Chemically stable (ph -) Mechanically stable Withstands repeated cleaning with base, and use of organic solvents, denaturants and surfactants Use same column for most biopolymers Fast QC analysis and process monitoring Reduced solvent consumption and analysis time Tosoh Bioscience offers a broad line of high efficiency columns for analysis and isolation of biomolecules by anion and cation exchange chromatography. In either mode of Ion Exchange Chromatography, the product line contains methacrylate-based as well as silica-based columns. Proteins, peptides, DNA and RNA derived oligonucleotides and other nucleic acid fragments are typical samples that are analyzed or isolated on TSK-GEL ion exchange columns. Silica-Based Ion Exchange Most suitable for analyzing smaller MW samples such as nucleotides, drug candidates, catecholamines and small peptides or proteins The methacrylate chemistry also forms the backbone of nonporous resin (NPR) columns such as DEAE-NPR, SP-NPR and DNA-NPR. These.6 mm ID columns are packed with.5 µm particles. High column efficiency coupled to low sample capacity restricts the application of these columns to fast analysis and micro-scale preparative isolation. TSK-GEL Anion Exchange Columns TSKgel Matrix* Particle Pore Functional Counter Exclusion Capacity Small Ion pk a Column Column Size Size Group Ion Limit, PEG** (mg BSA/mL) capacity hard- (µm) (Å) (Da) (meq/ml) ware*** BioAssist Q pma 0, ~000 Polyamine Cl - >5,000,000 70 0. 9. PEEK SuperQ-5PW pma 0, 000 Trimethyl-amino Cl -,000,000 00 >0.5. S, G DEAE-5PW pma 0,, 0 000 DEAE Cl -,000,000 0 0..5 S, G DEAE-NPR pma.5 ~ 0 DEAE Cl - 500 5 >0.5. S DNA-NPR pma.5 ~ 0 DEAE ClO - 500 5 >0.5. S DEAE-SW Silica 5 5 DEAE H PO - 0,000 ND >0.. S DEAE-SW Silica 0 50 DEAE Cl - 0,000 ND >0.. S QAE-SW Silica 0 5 Trimethyl-amino H PO - 0,000 ND >0.. S Sugar AXI PS-DVB 8 60 Trimethyl-amino HBO - ND >. S Sugar AXG PS-DVB 0 60 Trimethyl-amino HBO - ND >. S SAX PS-DVB 5 60 Trimethyl-amino Cl - ND >.0 S TSK-GEL Cation Exchange Columns TSK gel Matrix* Particle Pore Functional Counter Exclusion Capacity Small Ion pk a Column Column Size Size Group Ion Limit, PEG** (mg BSA/mL) capacity hard- (µm) (Å) (Da) (meq/ml) ware*** BioAssist S pma 7, ~00 Sulfopropyl Na+ ~,000,000 70 () 0.. PEEK SP-5PW pma 0,,0 000 Sulfopropyl Na+,000,000 0 () >0.. S, G CM-5PW pma 0, 000 Carboxymethyl Na+,000,000 5 () >0.. S, G SP-SW Silica 5 5 Sulfopropyl Na+ 0,000 ND 0.. S SP-NPR pma.5 ~ 0 Sulfopropyl Na+ 500 5 () >0.. S CM-SW Silica 5 5 Carboxymethyl Na+ 0,000 0 () >0.. S CM-SW Silica 0 50 Carboxymethyl Na+ 0,000 ND >0.. S SCX PS-DVB 5 60 Sulfonic acid Na+,H+ ND >.5 S OApak-A pma 5 000 Carbonic acid H+ ND >.5 S * pma = poly methacrylate; PS-DVB = polystyrene-divinylbenzene () γ-globulin; () hemoglobin ** Polyethylene glycol *** PEEK = polyetheretherketone, S = stainless steel, G = glass Phenomenex 57
Ion Exchange Chromatography (cont'd) Nucleotides Hemoglobin A C Column: TSKgel DEAE-SW Dimensions:.6 mm x 5 cm Part No.: CHO-807 Mobile Phase: A:Acetonitrile in 0. M phosphate, ph.0, 0/80 B:Acetonitrile in 0.5 M phosphate, ph.0, 0/80 Gradient: 0 min linear gradient from buffer A to B Flow Rate:.0 ml/min Detection: UV @ 60 nm Sample: Hemoglobin Standard. AMP. ADP. IMP 5. ATP. GMP 5 Column: TSKgel SP-NPR Dimensions:.6 mm x.5 cm Part No.: CHO-809 Mobile Phase: A:0.0 M MES, and 0.0 M HEPES-NaOH, ph 6.0 B:0.0 M MES, and 0.0 M HEPES-NaOH, ph 8.0 Gradient: 0 min linear gradient from % to 75 % buffer B (ph 6.66 to ph 7.) Flow Rate:.5 ml/min Detection: VIS @ 5 nm Sample: Hemoglobin Standard hemoglobin A C 0 min 0 6 8 0 min Anion Exchange PEEK Column I I I I I CH0-85 9685 BioAssist Q, 000 Å.6 5.0 0 CH0-807 0 BioAssist Q, 000 Å 0.0 0.0 Stainless Steel Columns: Polymer-based CH0-808 075 DEAE-NPR, nonporous.6.5.5 CH0-809 89 DNA-NPR, nonporous.6 7.5.5 CH0-80 8757 DEAE-5PW, 000 Å.0 7.5 0 CH0-076 076 DEAE-5PW, 000 Å 7.5 7.5 0 CH0-80 857 SuperQ-5PW, 000 Å 7.5 7.5 0 CH0-80 0869 Sugar AXI, 60 Å.6 5.0 8 CH0-80 0860 Sugar AXG, 60 Å.6 5.0 0 CH0-805 0757 SAX 6.0 5.0 5 Stainless Steel Columns: Silica-based CH0-806 0766 QAE-SW, 5 Å.6 5.0 5 CH0-807 0768 DEAE-SW, 5 Å.6 5.0 5 CH0-808 076 DEAE-SW, 50 Å 7.5 7.5 0 CH0-809 7088 DEAE-NPR Guard column.6 0.5 5 CH0-800 85 DNA-NPR Guard column.6 0.5 5 CH0-80 888 SuperQ-5PW Guardgel Kit 0 CH0-80 070 DEAE-5PW Guardgel Kit 0 CH0-80 5 DEAE-5PW Guard cartridge.0.0 0 CH0-80 0768 DEAE-SW Guardgel Kit 0 CH0-8 908 Guard cartridge holder.0.5 Cation Exchange PEEK Column I I I I I CH0-805 9686 BioAssist S, 00 Å.6 5.0 7 CH0-806 BioAssist S, 00 Å 0.0 0.0 Stainless Steel Columns: Polymer-based CH0-807 068 CM-5PW, 000 Å 7.5 7.5 0 CH0-808 8758 SP-5PW, 000 Å.0 7.5 0 CH0-078 076 SP-5PW, 000 Å 7.5 7.5 0 CH0-809 076 SP-NPR, nonporous.6.5.5 CH0-800 0756 SCX (Na+) 6.0 5.0 5 CH0-80 0758 SCX (H+) 7.8 0.0 5 CH0-80 665 OApak-A, 000 Å 7.8 0.0 5 Stainless Steel Columns: Silica-based CH0-80 0765 SP-SW, 5 Å.6 5.0 5 CH0-80 0767 CM-SW, 5 Å.6 5.0 5 CH0-805 076 CM-SW, 50 Å 7.5 7.5 0 CH0-806 069 CM-5PW Guardgel Kit 0 CH0-807 609 CM-5PW Prep Guardgel Kit 0 CH0-808 07 SP-5PW Guardgel Kit 0 CH0-809 5 SP-5PW Guard cartridge.0.0 0 CH0-8050 08807 SP-5PW Guardgel Kit, Glass 0 CH0-805 67 SP-5PW Guard column, Glass 0.0.0 CH0-805 609 SP-5PW Prep Guardgel Kit 0 CH0-805 079 SP-5PW Guard column 5.0 5.0 0 CH0-805 07650 CM-SW Guardgel Kit 0 CH0-8055 665 OApak-P Guard column 6.0.0 0 CH0-8 908 Guard cartridge holder.0.5 58 Phenomenex
Hydrophobic Interaction Chromatography - HIC TSK-GEL Hydrophobic Interaction Columns TSKgel Ether-5PW, TSKgel Phenyl-5PW, TSKgel Butyl-NPR TSKgel Ether-5PW and Phenyl-5PW columns now available in a mm ID format Large 000 Å pore size of the base matrix accommodates proteins up to 5 x 0 6 Da Polymeric resin is chemically and physically stable to changes in ph and ionic strength Compatible with a variety of organic solvents Hydrophobic Interaction Chromatography (HIC) is a separation technique based on the interaction between hydrophobic groups on a protein and a hydrophobic ligand on the solid support. HIC offers a distinct advantage for unstable proteins: it can be run using moderate concentrations of ammonium sulfate, which favors the stability of many proteins. Benefits of HIC HIC adds flexibility during method development as an alternative to Ion Exchange Chromatography Wide ph range (-) of the base resin enables robust cleaning options Seamless scalability from analytical to preparative scale Eliminates undesirable interactions with column hardware Dependence of Peak Width on Sample Loading in the Separation of Proteins Peak Width (ml).5.0.5.0 0..0 Sample Loading (mg) Column: TSKgel Phenyl-5PW Dimensions: 7.5 mm x 7.5 cm Part No.: CH0-806 Mobile Phase: 60 min linear gradient of (NH ) SO from.5 M to 0.0 M in 0. M phosphate buffer (ph 7.0) Flow Rate: 0.5 ml/min Temperature: 5 C Detection: UV @ 80 nm Sample:. Myoglobin. Ribonuclease A. Ovalbumin. a-chymotrypsin; concentration: 0.05 % to.6 % Hydrophobic Interaction Stainless Steel Columns I I I I I CH0-8056 8760 Ether-5PW, 000 Å.0 7.5 0.0 CH0-8057 086 Ether-5PW, 000 Å 7.5 7.5 0.0 CH0-8058 086 Ether-5PW, 000 Å.5 5.0.0 CH0-8059 655 Ether-5PW, 000 Å 55.0 0.0 0.0 CH0-8060 8759 Phenyl-5PW, 000 Å.0 7.5 0.0 CH0-806 0757 Phenyl-5PW, 000 Å 7.5 7.5 0.0 CH0-806 07656 Phenyl-5PW, 000 Å.5 5.0.0 CH0-806 0798 Phenyl-5PW, 000 Å 55.0 0.0 0.0 CH0-806 97 Butyl-NPR, nonporous.6.5.5 PEEK Columns CH0-8065 00 BioAssist Phenyl-5PW, 7.8 5 0.0 000 Å CH0-8066 56 Ether-5PW Guard cartridge.0.0 0.0 CH0-8067 t 086 Ether-5PW Guardgel Kit 0.0 CH0-8068 55 Phenyl-5PW Guard cartridge.0.0 0.0 CH0-8069** 0765 Phenyl-5PW Guardgel Kit 0.0 CH0-8070 0796 Phenyl-5PW Guard column 5.0 5.0 0.0 CH0-8* 908 Guard cartridge holder.0 * For.0 mm ID cartridges ** For 7.5 mm ID columns t For.5 mm ID columns For 55 mm ID columns Phenomenex 59
Tosoh Bioscience llc Affinity Chromatography TSK-GEL Affinity Columns TSKgel ABA-5PW, TSKgel Boronate-5PW, TSKgel Chelate-5PW, TSKgel Tresyl-5PW Enhanced access of large proteins to affinity ligands High efficiency and wide ph range (-) The Tosoh Bioscience TSK-GEL Affinity Chromatography (AFC) column line consists of three group-specific stationary phases and one activated packing material called Tresyl-5PW. Affinity chromatography offers the highest level of specificity and selectivity in biomolecular separations and purifications. Characteristics of TSK-GEL AFC Columns Column Packing Ligand Type Ligand Concentration Adsorption Capacity Sample ABA-5PW p-aminobenzamidine not available -mg/ml - resin trypsin Boronate-5PW m-aminophenyl boronate not available 0 µmole/ml - resin sorbitol Chelate-5PW iminodiacetic acid 0 µmole/ml - resin not available not available Tresyl-5PW tresyl ca. 0 µmole/ml - resin >60 mg/g dry resin soybean (coupling capacity) trypsin inhibitor Affinity Stainless Steel Columns I I I I I CH0-807 067 ABA-5PW, 000 Å 7.5 7.5 0 CH0-807 066 Boronate -5PW, 000 Å 7.5 7.5 0 CH0-807 0865 Chelate-5PW, 000 Å 7.5 7.5 0 CH0-807 56 Tresyl-5PW, 000 Å 7.5 7.5 0 PEEK Columns CH0-8075 00 BioAssist Chelate, 000 Å 7.8 5 0 CH0-8076 7 ABA-5PW Guardgel Kit CH0-8077 5 Boronate-5PW Guardgel Kit CH0-8078 0867 Chelate-5PW Guardgel Kit Bulk Packing CH0-8079 608 Tresyl-5PW, g dry gel* 0 * g is approximately.5 ml 60 Phenomenex
ToyoScreen Process Method Development Columns Historically, resin screening was accomplished by packing various bulk resins into small columns by hand, requiring significant investments in time and cost. In order to improve the efficiency of resin screening experiments, pre-packed ToyoScreen columns, containing the most popular Toyopearl resins, now provide a low cost, convenient way to evaluate different Toyopearl ligands. The ToyoScreen Series are for ion exchange, hydrophobic interaction and affinity chromatography. The system consists of a set of six ml or 5 ml cartridge columns to be mounted in a holder. The columns exhibit reproducibility and show high numbers of theoretical plates. The columns are compatible to most common and FPLC systems and are reusable, as is the holder. Part No. Mfr. No. Description Unit Price Ion Exchange I I I CH0-8080 60 ToyoScreen DEAE-650M ml x 6 each CH0-808 6 ToyoScreen DEAE-650M 5 ml x 6 each CH0-808 6 ToyoScreen SuperQ-650M ml x 6 each CH0-808 6 ToyoScreen SuperQ-650M 5 ml x 6 each CH0-808 6 ToyoScreen QAE-550C ml x 6 each CH0-8085 65 ToyoScreen QAE-550C 5 ml x 6 each CH0-8086 66 ToyoScreen CM-650M ml x 6 each CH0-8087 67 ToyoScreen CM-650M 5 ml x 6 each CH0-8088 68 ToyoScreen SP-650M ml x 6 each CH0-8089 69 ToyoScreen SP-650M 5 ml x 6 each CH0-8090 70 ToyoScreen SP-550C ml x 6 each CH0-809 7 ToyoScreen SP-550C 5 ml x 6 each Hydrophobic Interaction CH0-809 7 ToyoScreen Ether-650M ml x 6 each CH0-809 7 ToyoScreen Ether-650M 5 ml x 6 each CH0-809 7 ToyoScreen Phenyl-650M ml x 6 each CH0-8095 75 ToyoScreen Phenyl-650M 5 ml x 6 each CH0-8096 76 ToyoScreen Butyl-650M ml x 6 each CH0-8097 77 ToyoScreen Butyl-650M 5 ml x 6 each CH0-8098 78 ToyoScreen Hexyl-650C ml x 6 each CH0-8099 79 ToyoScreen Hexyl-650C 5 ml x 6 each CH0-800 80 ToyoScreen PPG-600M ml x 6 each CH0-80 8 ToyoScreen PPG-600M 5 ml x 6 each CH0-80 8 ToyoScreen SuperButyl-550C ml x 6 each CH0-80 8 ToyoScreen SuperButyl-550C 5 ml x 6 each Part No. Mfr. No. Description Unit Price Affinity I I I CH0-80 8 ToyoScreen AF-Chelate-650M ml x 6 each CH0-805 85 ToyoScreen AF-Chelate-650M 5 ml x 6 each CH0-806 86 ToyoScreen AF-Blue HC-650M ml x 6 each CH0-807 87 ToyoScreen AF-Blue HC-650M 5 ml x 6 each CH0-808 88 ToyoScreen AF-Red-650M ml x 6 each CH0-809 89 ToyoScreen AF-Red-650M 5 ml x 6 each Mixed Packs CH0-80 9 ToyoScreen IEC Anion-650M ml x Grades x each CH0-8 9 ToyoScreen IEC Anion-650M 5 ml x Grades x each CH0-8 9 ToyoScreen IEC Cation-650M ml x Grades x each CH0-8 95 ToyoScreen IEC Cation-650M 5 ml x Grades x each CH0-8 96 ToyoScreen IEC-650M ml x 6 Grades x each CH0-85 97 ToyoScreen IEC-650M 5 ml x 6 Grades x each CH0-86 98 ToyoScreen HIC-650M ml x 6 Grades x each CH0-87 99 ToyoScreen HIC-650M 5 ml x 6 Grades x each ToyoScreen Column Accessories CH0-88 00 ToyoScreen Column Holder ea CH0-89 9 ToyoScreen Holder w/ Fittings Includes Mfr. Nos. 00, 95, and 96 CH0-80 95 Column to Column Connector ea AQ0-8 96 M6 Adaptor ea ToyoScreen, TSK-GEL, TSKgel, Tosoh Bioscience, Toyopearl MegaCap and BioAssist are registered trademarks of Tosoh Corporation, the parent company of Tosoh Bioscience. Phenomenex 6