Gel Filtration. Sorbadex - For process purification of proteins and nucleic acids

Transcription

1 This is a separation method for purifying interesting relatively high molecular weight substances such as proteins, glycoproteins, lipoproteins, and the like from lower molecular weight substances such as buffer constituents, salts, cell debris, and the like. The driving force for movement through the adsorbent can be A. Gravity whether through classic glass columns or Polypropylene columns B. Pressure (Flash Chromatography and FPLC) or C. Centrifugal force (Spin Columns and 96 Well Plates). Sorbadex - For process purification of proteins and nucleic acids SorbaRes - For Separation of Proteins, Peptides, Nucleic acids, and other Biomolecules Clarion P - Hydrated gel filtration columns for protein purification, desalting, and buffer exchange Clarion N - Hydrated gel filtration columns for nucleic acid purification and desalting Sorbasep - For desalting, removal of small molecules, and buffer exchange using liquid chromatography systems Clarion Plates - For desalting, buffer exchange, and removal of free labels FPLC Columns - Laboratory glass columns 52

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3 - Size Exclusion Chromatography Sorbadex - For process purification of proteins and nucleic acids Sorbadex is a beaded composite material composed partially of cross linked dextran. It exhibits high selectivity, high resolution and chemical stability. Sorbadex is a size exclusion matrix. Molecules purified with Sorbadex are separated according to size. Smaller molecules pass significantly slower through the column than larger molecules. Buffer and ph effects on resolution are minimal. The size exclusion cut-off for Sorbadex -25 is set at 5 kd for proteins and 10 bp for nucleic acids. For Sorbadex -50, the cut-offs are 25 kd and 20 bp. Purified biomolecules are not significantly diluted when processed using Sorbadex. Sorbadex is autoclavable at 121 C, ph 7 for 30 minutes and is stable in all commonly used buffers, including: 0.2M NaOH; 0.2M HCl; 1M acetic acid; 8M urea; 6M guanidine HCI; 1% SOS, 24% Ethanol; 30% Propanol; and 30% Acetonitrile. Sorbadex bulk powders are available in 100g, 500g, 1kg, 5kg, 10kg, and 100kg package sizes. Cat. No. Product Fractionation range (globular proteins) -Mr Sorbadex -25 superfine Sorbadex -25 fine Sorbadex -25 medium Sorbadex -50 superfine Sorbadex -50 fine Sorbadex -50 medium ph stability Bed volume ml/g dry Sorbadex Bead size (dry) 1-5 kda2 to µm 1-5 kda2 to µm 1-5 kda2 to µm 1-30 kda2 to µm 1-30 kda2 to µm 1-30 kda2 to µm Sorbadex -50 fine Hydrated in phosphate buffered saline; ph 7.4, Sorbadex -25 medium with 0.2% sodium azide Sorbadex 25 fine Lipophilic Hydrophilic Matrix, hydroxypropylated 54

4 Sorbadex High Performance Results Sample: Mouse IgG in PBS Conc.: 12.5 mg/ml Volume: 4500 ml Total IgG: grams Buffer: 100 mm Glycine-HCl, ph 2.0 neutralised with 1 M Tris-HCl ph 8.0 System: ÄktaPilot (GE Healthcare) Column: Sorbadex-25 M in BPG 300/500 Diameter: 30 cm Gel Volume: 25 Liters 3500 Pressure: 1.0 bar (0.1 mpa or 13 psi) 50 Flow rate during sample loading: 400 ml 3000 Flow rate during equilibation and elution: 500 ml ma U 40 Total run time: 63 min 2500 ms/cm ma U ms/cm For laboratory use only, not for drug, household or other use. Specifications Cat. No Product Description Fractionation range (globular proteins) - Mr Fractionation range (dextrans) - Mr Sorbadex -25 Matrix Dry Beads Superfine Fine Medium Medium - in Phosphate Buffered Saline, ph 7.4 with 0.2% soduim azide Sorbadex -50 Matrix Dry Beads Superfine Fine Medium 1-5 kd 1-30 kd kd kd Bead structure Cross-linked dextran composite Cross-linked dextran composite Bead size (Dry) 20-50µm 20-80µm µm µm 20-50µm 20-80µm µm Bead size (Wet) 35-90µm µm µm µm µm µm µm Obeys Darcy's Law Obeys Darcy's Law Obeys Darcy's Law Chemical stability All commonly used buffers, including: 0.2 M Na0H; 0.2M HCI; 1M acetic acid; 8M urea; 6M guanidine HCI; 1% SDS, 24% Ethanol; 30% Propanol; 30% Acetonitrile. All commonly used buffers, including: 0.2 M Na0H; 0.2M HCI; 1M acetic acid; 8M urea; 6M guanidine HCI; 1% SDS, 24% Ethanol; 30% Propanol; 30% Acetonitrile. ph stability 2.0 to to 13.0 Autoclavable t 121 C, ph 7 for 30 minutes t 121 C, ph 7 for 30 minutes v(ml) 55

5 Sorbadex Matrix: For process purification of proteins and Nucleic Acids High Performance Results Protein Desalting from 0.8M NaCl IgG (280nm): dark blue line NaCl (µs): gold line 1 mg IgG anti-rabbit in 1 ml 0.8 M sodium chloride Water elution Removal of FITC from IgG IgG (280nm): dark blue line FITC (550nm): gold line 1 mg IgG anti-rabbit and 0.1 µmol FITC in 1mL DMSO/ NaHCO3 PBS Elution Oligo Desalting from 0.8M NaCl Oligo (260nm): dark blue line NaCl (µs): gold line 1 mg oligonucleotide (18-mer) in 1 ml 0.8 M NaCl Removal of Rhodamine after labeling Oligo (260nm): dark blue line TAMRA (550nm): gold line 1 mg oligonucleotide (18-mer) and 0.5 µmol TAMRA in 1 ml DMSO/NaHCO3 Removal of Ammonia (33%) Dextran Blue (260nm): dark blue line NH3 (ph): gold line 0.5 mg Dextran Blue (Mr 2,000,000) in 1 ml ammonia (33%). Water elution. Removal of Ammonia (33%) from Oligo Oligo (260nm): dark blue line NH3 (ph): gold line 1 mg oligonucleotide (18-mer) in 1 ml ammonia (33%) after cleavage. 56

6 Sorbadex Matrix: For process purification of proteins and Nucleic Acids Hydration, Filling and Packing Columns Sorbadex can be hydrated in aqueous media of choice containing no more than 20% alcohol. Time of Hydration should be a minimum of 3 hours at room temperature or 1 hour at 90 C. Hydrated Sorbadex is provided in a settled gel volume to buffer volume ratio of 1:1, degassed and ready to use. Filling a column requires that the slurry be not too thick as to retain air bubbles. A settled gel volume to buffer volume ratio of 3:1 is optimal for this purpose. The remaining buffer may be used later for column packing. To fill, pour the slurry into a tilted column. Alternatively, a funnel may be used, with the tip of the funnel touching the inside wall of the column to prevent splashing of the slurry. A gel reservoir or column extension is desirable for filling the whole column in a single operation. To finish packing the gel bed, a peristaltic pump may be employed. Use as high a flow rate as possible without deforming the beads. Sorbadex can be pressurized up to 3 bar. Precautions for Safe Handling As part of good industrial and personal hygiene and safety procedure, avoid all unnecessary exposure to the chemical substance and ensure prompt removal fron skin, eyes and clothing. Keep container tightly closed. Suitable for any general chemical storage area. Containers of this material may be hazardous when empty since they retain product residues (dust, solids); observe all warning and precautions listed for the product. Maximum operating pressure: Generally obeys Darcy s Law: U=K º P L where U = linear flow rate, cm/hour P = pressure drop over gel bed, cm H 2 0 L = bed ht, in cm K º = 9 for S-25 Superfine, 30 for S-25 Fine, 80 for S-25 Medium, 13.5 for S-50 Superfine 36 for S-50 Fine, 145 for S-50 Medium 57

7 SorbaRes IEC - For process purification of proteins and nucleic acids SorbaRes Resins are specially designed for the chromatographic separation of biomolecules. Based on a highly porous hydrophilic polymethacrylate matrix, the rigid, spherical, uniform particle size enables high speed chromatographic operation with high production efficiency. SorbaRes S SorbaRes Q SorbaRes CM SorbaRes DA Matrix Crosslinked polymethacrylate Type Strong Cation Strong Anion Weak Cation Weak Anion Particle Size 30μm and 60μm Ion exchange group -SO 3 - -N(CH 3 ) 3 -COOH -N(CH 3 ) 2 ph Range Regular use: 3 12 Short term use: 1 13 Temp. range 2-45 C Shipping solvent Static binding capacity* (g/l-resin) Ion exchange capacity (meq/ ml-resin) 20 mm sodium phosphate in 20% ethanol min. 100 min. 100 min. 90 min. 77 min min min min. 0.1 Cat. # Description Qty L SorbaRes S30 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Sulfonate, Strong Cation Type, 30µm 25ml L SorbaRes S60 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Sulfonate, Strong Cation Type, 60µm 25ml L SorbaRes Q30 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Quat. Ammonium, Strong Anion Type, 30µm 25ml L SorbaRes Q60 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Quat. Ammonium, Strong Anion Type, 60µm 25ml L SorbaRes CM30 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Carboxymethyl, Weak Cation Type, 30µm 25ml L SorbaRes CM60 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Carboxymethyl, Weak Cation Type, 60µm 25ml L SorbaRes DA30 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Dimethylamine, Weak Anion Type, 30µm 25ml L SorbaRes DA60 Ion Exchange Resin, Crosslinked Polyhydroxymethacrylate, Dimethylamine, Weak Anion Type, 60µm 25ml For more selections and sizes, visit our website or contact your personal Product Specialist. SorbaRes SEC - For process purification of proteins and nucleic acids SorbaRes Size Exclusion Resins are based on a cross linked polymethacrylate resin that has been treated with a hydrophillic layer. SorbaRes Size Exclusion Resins offer increased strength and flow rate. No salts are required; results are achieved with water only. Cat. # Description Qty SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 120Å 10g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 120Å 25g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 120Å 50g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 200Å 10g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 200Å 25g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 200Å 50g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 600Å 10g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 600Å 25g SorbaRes Size Exclusion Media, Highly Crosslinked Polyhydroxymethacrylate, 10µm, 600Å 50g For more selections and sizes, visit our website or contact your personal Product Specialist. 58

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9 Clarion P Hydrated gel filtration columns for protein purification, desalting, and buffer exchange Clarion P Columns are specifically designed for rapid and efficient removal of small molecultes (salts, dyes, ammonia, haptens, biotin, etc.) from anibodies, enzymes, and other proteins. Ultrapure gel and specially treated sinter frits ensure outstanding reolution and high selectivity. The gel matrix of Clarion is Sorbadex -25, a beaded composite material coprised of ultrapure cross-linked dextran. It exhibits high selectivity, high resolution and chemical stability. Molecules puridies with Sorbadex -25 are separated according to size. Smaller molecules pass significantly slower through the column than larger molecules. Buffer and ph effects on resolution are minimal. The molecular weight cut-off (MWCO) for Sorbadex -25 is 5 kd for proteins. Proteins larger than 5 kd are typically purified with a 1.5-fold elution volume. Absopr tion Absopr tion Conductivity Elution volume (ml) Removal of fluorescent dye Oval bumin (280 nm): dark blue line FAM (490 nm): gold line Elution profile overlay of albumin (5 mg OvA) and free dye (2.5 µmol FAM) in DMSO/NaHCO 3, elution with water (5.0 ml sample volume) Elution volume (ml) Desalting of protein solution (1 mg anti-rabbit IgG in 1 ml o.8 M NaCI), elution with water (dark blue line: protein-280 nm; gold line: salt-µs/cm.) 7. 0 Cat. # Name Sample Volume Pack Size Clarion P µl 50 Columns Clarion P50 0.5ml 50 Columns Clarion P ml 50 Columns Clarion P ml 25 Columns Clarion P ml 10 Columns Clarion P ml 10 Columns Clarion P ml 1 Column 60

10 Desalt Hydrated with pure, deionized water. Pre-swollen, pre-packed and ready-to-use. Clarion MINI Columns are used for quick and efficient desalting, buffer exchange and/or removal of dyes and small molecules from proteins (S-25 greater than 5 kd, S-50 greater than 25 kd). Purified proteins are eluted into pure, deionized water (Caution! Some proteins may precipitate in pure water with low ionic strength!) The columns are sterile packed, pre-swollen and ready-to-use. Desalt AZ Hydrated with pure deionized water and stabilized with 0.02% Sodium azide. Pre-swollen, pre-packed and ready-to-use. Clarion MINI Columns are used for quick and efficient desalting, buffer exchange and/or removal of dyes and small molecules from proteins greater than 5 kd. Purified proteins are eluted into pure, deionized water (Caution! Some proteins may precipitate in pure water with low ionic strength!) The columns are stabilized with 0.02% sodium azide, pre-swollen and ready-to-use. Desalt Hydrated with 1 mm TRIS ph 6. Pre-swollen, pre-packed and ready-to-use. Clarion TRIS MINI Columns are used for rapid buffer exchange and/or removal of dyes and small molecules from proteins (S-25 greater than 5 kd, S-50 greater than 25kD). Purified proteins are eluted into 1 mm TRIS, ph 6. The columns are sterile packed, pre-swollen and ready-to-use. TRIS Hydrated with 1 mm TRIS ph 6. Pre-swollen, pre-packed and ready-to-use. Clarion TRIS MINI Columns are used for rapid buffer exchange and/or removal of dyes and small molecules from proteins (S-25 greater than 5 kd, S-50 greater than 25kD). Purified proteins are eluted into 1 mm TRIS, ph 6. The columns are sterile packed, pre-swollen and ready-to-use. PBS Hydrated with Phosphate Buffered Saline ph 7. Pre-swollen, pre-packed and ready-to-use. Clarion Clarion PBS MINI Columns are used for rapid buffer exchange and/or removal of dyes and small molecules from proteins (S-25 greater than 5 kd, S-50 greater than 25 kd). Purified proteins are eluted into Phosphate Buffered Saline (PBS, ph 7). The columns are sterile packed, pre-swollen and ready-to-use. Easy 4 Step Protocol 1. Column Preparation Remove the cap from the top and then the white bottom cap of the Clarion P or Clarion N Column. Allow excess column fluid to drain (via gravity) into a suitable waste reservoir. 2. Column Equilibration Equilibrate the column by loading it with 5x the bed volume of water or buffer (use the same buffer for equilibration and elution). Allow the equilibration buffer to drain completely. 3. Sample Application Transfer the sample to the Clarion P or Clarion N Column. Allow the sample to enter the gel completely. 4. Elution Place a tube for sample collection under the Clarion P or Clarion N Column. Trander the elution buffer to the column and elute the purified sample. 61

11 Clarion N Hydrated gel filtration columns for nucleic acid purification and desalting Clarion N Columns are specifically designed for rapid and efficient removal of small molecules (salts, dyes, ammonia, haptens, biotin, etc.) from nucleic acids. Ultrapure gel and specially treated sinter frits ensure outstanding resolution and high selectivity. The gel matrix of Clarion is Sorbadex -25, a beaded composite materula comprised of ultrapure cross-linked dextran. It exhibits high selectivity, hight resolution and chemical stability. Molecules purified with Sorbadex -25 are separated according to size. Smaller molecules pass sugnificantly slower through the column than larger molecules. Buffer and ph effects on resolution are minimal. The molecular weight cut-off (MWCO) for Sorbadex -25 is 10 bases for nucleic acids. Oligonucleotides larger than 10 bases are typically purified with 1.5-fold elution volume. Absopr tion ph Absopr tion Elution volume (ml) Separation of 10 M ammonia and olugonucleotide after cleavage from solid support and removal of protecting groups (18-mer, Scale: 0.2 µmol, 1 ml sample volume). Elution with water Elution volume (ml) Elution profile overlay of 2.5 µmol 5-TAMRA and 0.25 µmololigonucleotide (2.5 ml sample volume). SEQ S-50 Hydrated with pure, deionized water. Pre-swollen, pre-packed and ready-to-use. Clarion SEQ S-50 MINI Columns are used for rapid removal of dyes and dideoxy terminators from sequencing reactions and for desalting of oligonucleotides greater than 20 base pairs. Purified nucleic acids are eluted into pure, deionized water. The columns are sterile packed, pre-swollen and ready-to-use. Desalt S-25N Hydrated with pure, deionized water. Pre-swollen, pre-packed and ready-to-use. Clarion Desalt S-25N MINI Columns are used for rapid removal of salts, dyes and other small molecules from nucleic acids larger than 10 base pairs. Purified nucleic acids are eluted into pure, deionized water. The columns are sterile packed, pre-swollen and ready-to-use. Cat. # Name Sample Volume Pack Size Clarion N µl 50 Columns Clarion N50 0.5ml 50 Columns Clarion N ml 50 Columns Clarion N ml 25 Columns Clarion N ml 10 Columns Clarion N ml 10 Columns Clarion N ml 1 Column 62

12 Sorbasep - FPLC Desalting Columns For desalting, removal of small molecules, and buffer exchange using liquid chromatography systems SorbaSep FPLC Desalting Colimns are designed for: Separating larger biomolecules (i.e. proteins such as antibodies, enzymes, or larger nucleic acids) from unwanted smaller molecules. Buffer exchange, desalting, removal of low molecular weight contaminants, and reaction terminations. Simple, rapid and reproducible separation using a syringe pump or liquid chromatography system. The fractionation range for globular proteins is between 1 and 5 kd. The molecular weight cut-off (MWCO) is approximately 5 kd, which ensures efficient separation of proteins / peptides / biomolecules larger than 5 kd from lower molecular weight of less than 1 kd. SorbaSep Desalting FPLC Columns contain Sorbadex -25 Superfine, a beaded composite size exclusion matrix. It exhibits high flow rates, excellent resolution and chemical stability. Buffer and ph effects on resolution are minimal. High Performance Results 2ml/mi n FAM* Sample: 1 ml of 2 mg/ml BSA & 100 µm of in PBS ph 7.4 (0.05% NaN3) Flow rate: 2 ml/min. Eluent: PBSpH 7.4 (0.05% NaN3) Detection: Abs. at 280 nm and 490 nm Absopr tion 2ml/mi n BSA Time (Minutes) Column Bed Volume Size of Eluted Proteins System Compatibility Column Dimensions Column Body material Column Portsl Support Bead size Maximum Back Pressure3 Recommended Flow Rate1 Maximum Recommended Flow Rate Storage Temperature Storage Solution 5ml >5 kd Specifications Automated liquid chromatography systems (MPLC, FPLC, AKTA, ect.) Peristalic Pump Syringe 1.6 cm inner diameter x 2.5 cm height Polypropylene Inlet (1/16') Female Outlet (1/16') male Sorbadex -25 Superfine µm (hydrated) bar (0.3 MPa) to 5 ml/min 10 ml/min Ambient 20% (v/v) ethanol Cat. # Description Pack Size SorbaSep FPLC Desalting 5 x 5 ml Columns SorbaSep FPLC Desalting 100 x 5 ml Columns 63

13 Clarion - Plates For desalting, buffer exchange, and removal of free labels Clarion N Plates are precision filled with Sorbadex a beaded composite material comprised of ultrapure cross-linked dextran. They are specifically designed for desalting, buffer exchange and rapid removal of compounds such as dye terminators, dntps, salts, nucleic acid fragments, biotin and other low molecular weight haptens. Sorbadex exhibits high selectivity, high resolution and chemical stability. The molecular weight cut-off (MWCO) for Sorbadex -25 is 5 kd for proteins and 10 bases for nucleic acids. For Sorbadex -50, the cut-offs are 25kD and 20 bases respectively. Standard Clarion Plates are hydrated with sterile purified water without preservatives, salts or buffers. The filtration plate is made from sterile medical-grade polypropylene. Each gel bed is supported on an individual ultra high molecular weight PE filter membrane with pore size of 25 microns. Products may be collected into standard 96 or 384 well format collection plates (not supplied) for subsequent processing. The purification protocol consists of a simple 2 to 3 minute spin to remove excess fluid from the wells. The samples are then loaded to the plate and spun again for two to three minutes to purify without significant dilution. Total hands on time is under 7 minutes. Cat. # Product # No. of Wells Clarion LD50 Plate 135 µl Well Volume Clarion SD50 Plate 400 µl Well Volume Clarion LD50 Plate 400 µl Well Volume Clarion SD50 Plate 800 µl Well Volume Clarion SD25 Plate 800 µl Well Volume Clarion 96 Plate 1000 µl Well Volume Clarion 96 Plate 2000 µl Well Volume Well Vol. Plate Height Max Sample Vol. 64 Gel Bed Vol. Matrix Short or long Drip Directors For Proteins / Nucleic Acids greater than Mode of Operation µL 15mm 10µL 100µl S-50SF Long 25 kd/20 bases Centrifuge 3 minutes at 1000xG µL 20mm 20µL 320µl S-50SF Short 25 kd/20 bases Centrifuge 3 minutes at 1000xG µL 20mm 20µL 320µl S-50SF Long 25 kd/20 bases Centrifuge 3 minutes at 1000xG µL 31mm 30µL 400µl S-50SF Short 25 kd/20 bases Centrifuge 3 minutes at 1000xG µL 31mm 30µL 400µl S-50SF Short 5 kd/10 bases Centrifuge 3 minutes at 1000xG µL 38mm 40µL 650µl S-50SF Long 25 kd/20 bases Centrifuge 3 minutes at 1000xG µL 44mm 80µL 1300µl S-50SF Long 25 kd/20 bases Light Vacuum

14 FPLC Columns SNAP - Laboratory Glass Columns "Next Generation" technology for high-performance preparative chromatography Liquid preparative chromatography is a widely used purification technique for a broad range of compounds, and for FPLC the common targeted molecules include proteins, peptides and nucleic acids. With the emergence of smaller particle, higher performance chromatography media, there has been an identified need for higher pressure column hardware that can handle the increased back pressure loading in a safe column configuration. Traditionally this has been addressed with stainless steel hardware, which does not allow the scientist visibility of the column contents. SNAP Laboratory Glass Columns are designed to address these evolving demands. The column technology exceeds what is currently available. Careful choice in materials of construction, combined with customer feedback, has driven the design so that biocompatibility can be achieved in virtually any application. Simple, yet innovative design ensures frustration-free results. Maximum Pressure Rating Column ID Pressure (bars) Pressure (PSI) Column Part Aqueous Buffer Version Solvent Resistant Version Body Precision Bore Glass Precision Bore Glass Pistons Acetyl PEEK O-Rings EPDM or Viton Viton or Kalrez Frit Polyethylene (5 µm or 10 µm) Stainless Steel or Teflon (2 µm or 10 µm) Temperature Range 4-40 C 4-40 C Height Adjustment Short/Short, Short/Long, Long/Long pistons Short/Short, Short/Long, Long/Long pistons Connections 1/4'-28 female screw thread 1/4'-28 female screw thread 65

15 SNAP Columns - Features and Benefits: 1. HIGHER PRESSURE RATINGS INCORPORATING GLASS CONSTRUCTION Pressures to 40 Bars (580 psig) Full view of bed unlike stainless steel Rugged construction for hard lab use 2. LINEAR MOTION OF PISTON Due to the true linear motion of piston there is NO scraping of the bed surface or induced TORSIONAL load imposed on the packed bed assuring true linear compression. 3. CALIBRATED GLASS TUBING Superior glass tolerance eliminates seal adjustment. True bore tubing reduces wall effect 4. TRUE FRITS Columns are supplied with true sintered material frits assuring good flow distribution and minimize dead volume. 5. ROBUST INLET AND OUTLET CONNECTIONS Connections are made externally and are visible if any leakage should occur. 6. FINE THREAD ADJUSTMENT Columns are provided with fine thread adjustment allowing better and more precise piston control. 7. DOUBLE PISTON ADJUSTMENT Columns are provided with piston adjustment from both ends allowing greater flexibility of bed adjustments. 8. QUICK RELEASE ENDS ELS (patented) column end closure design allows easy assembly and disassembly of columns even on large diameters. Inherently selflocking design ensures user safety. 9. GRADUATED GLASS All columns are supplied with column graduations on the glass exterior to make bed measurements quick and simple. 10. CONFIGURABLE Columns can be custom configured based on mobile phase conditions and meet the exact needs of the chromatographer. Packing Adaptor Since most columns will be wet or slurry packed, a packing adaptor should be used. SNAP column packing adaptors have a number of advantages. The first is that they are easy to install and remove. Secondly, the packing adaptors have a sufficient pressure rating, appropriate for the columns they are mounted on, allowing for adequate flow conditions. Third, they are the same diameter as the column they are mounted on avoiding issues with turbulent flow at the interface or joining point. All this adds up to ease of use, safe operations and good results! These consist of a coupling unit and glass body of same column ID as column to be packed. Packing Adaptor consists of: SNAP coupling unit (cast resin) with seal ring insert (PTFE) AB-Version with two Viton O-Rings SR-Version with two Kalrez or Viton O-Rings 66

16 Standard Available SNAP Configurations Col. ID Bed L. Press. Short/ Fixed (SF) Bed Length Short/ Fixed (SF) Volume Long/ Fixed (LF) Bed Length Long/ Fixed (LF) Volume Short/ Short (SS) Bed Length Short/ Short (SS) Volume Long/ Short (LS) Bed Length Long/ Short (LS) Volume Long/ Long (LL) Bed Length Long/ Long (LL) Volume With the Option of Customized Solutions Custom column lengths Customized material configurations Heating and cooling jackets Custom end connections Any unique specifications upon request Reduce overall cost by purchasing the next columns in cartridge form. Also, columns can be stored without rotating clamps. 67

17 FPLC Columns - Upscale and IsoKrom Upscale Small and mid size low pressure pilot / process columns from 90 mm to 450 mm diameter with bed lengths up to 900 mm. Upscale offers a glass column solution with superior flow distribution as well as a positive (no dead volume) mechanical seal. Design eliminates metal contact with wetted parts. HIGHLIGHTS: Simple Sanitary Design Low Dead Volume Reliable Mechanical Seal Low Maintenance Scalable Replaces BPG Cross-reference tables comparing Upscale to GE Healthcare (formerly Pharmacia) LC Columns. Sorbtech Cat. # ID Bed Height Volume Pressure Rating mm 0-25 mm L 6 bar mm mm L 6 bar mm mm L 6 bar N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A mm 0-25 mm L 4 bar mm mm L 4 bar mm mm L 4 bar mm 0-21 mm L 3 bar mm mm L 3 bar mm mm L 3 bar mm 0-21 mm L 2 bar mm mm L 2 bar mm mm L 2 bar GE Cat. # ID Bed Height Volume Pressure Rating BPG 100/ mm 0-26 mm 0-2 L 8 bar BPG 100/ mm mm L 8 bar BPG 100/ mm mm L 8 bar BPG 140/ mm 0-26 mm 0-4 L 6 bar BPG 140/ mm mm L 6 bar BPG 140/ mm mm L 6 bar BPG 200/ mm 0-26 mm L 6 bar BPG 200/ mm mm L 6 bar BPG 200/ mm mm L 6 bar BPG 300/ mm 0-26 mm L 4 bar BPG 300/ mm mm L 4 bar BPG 300/ mm mm L 4 bar BPG 450/ mm 3-23 mm L 2.5 bar BPG 450/ mm mm L 2.5 bar BPG 450/ mm mm L 2.5 bar Note: Frit Porosity is 20 μm for UpScale and 23 µm for GE IsoKrom Series These low and medium pressure fixed bed, side packed columns offer zero dead volume technology. Sizes are available from 95 mm to 2,000 mm with no practical limit on bed length. Simple construction uses standard components, providing low back-end cost of ownership. HIGHLIGHTS: All wetted parts are either autoclavable, disposable and/or can be completely taken apart for thorough cleaning. Simple & Robust - IsoKrom is designed around standard industrial parts, to provide a lower price and higher quality. Reduced dead-space by volume (V) and length (L). Fixed volume, packing through the wall. No metal parts contact the solution. 68

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