Prepacked chromatography columns for ÄKTA systems

Transcription

1 Prepacked chromatography columns for ÄKTA systems Selection guide gelifesciences.com

2 Select the chromatography technique of your choice below This guide gives you an overview of prepacked columns from GE Healthcare Life Sciences that can be used with ÄKTA protein purification systems. The columns are grouped by chromatography techniques. In each section, you will access product information and tips for product selection. Size exclusion chromatography, desalting/buffer exchange (SEC)...4 Ion exchange chromatography (IEX)...6 Affinity chromatography (AC) purification...8 Affinity chromatography (AC) antibody purification...10 Affinity chromatography (AC) purification of specific groups of biomolecules Hydrophobic interaction chromatography (HIC)...12 Multimodal chromatography (MMC)...13 Reversed phase chromatography (RPC)...13 Once you have selected your column, check its compatibility with your ÄKTA system on page 3. Optimize your results by combining chromatography techniques Significant improvements can be achieved by using a combination of different chromatography techniques. Examples of logical combinations of chromatography techniques that can be used for different types of proteins according to the principles of the proven capture, intermediate purification, and polishing (CiPP) purification strategy are shown below. Learn more from the Strategies for Protein Purification handbook, which can be downloaded at gelifesciences.com/proteinhandbooks Tagged protein Antibody Un Capture: Isolate, concentrate, stabilize AC AC IEX HIC Intermediate purification: Remove bulk impurities IEX* HIC* IEX* Polishing: Remove final contaminants SEC 1 SEC* SEC SEC Pure protein * These steps are not always used and will depend on the level of purity needed. Examples of logical combinations of chromatography techniques according to CiPP strategy. 2 KA SG

3 Check the compatibility of GE s column formats with your ÄKTA system type HiTrap HiScreen HiPrep HiLoad RESOURCE Tricorn Precision Main use Bed (ml) or column inner diameter (mm) Easy to use with a syringe, peristaltic pump, or chromatography system Optimized for method and process development Convenient scale up; preparative size exclusion chromatography Preparative purification with high resolution 1 ml 5 ml 4.7 ml 16 mm 26 mm 16 mm 26 mm 1 ml 3 ml Fast with good resolution High quality and high resolution Micropurification and analysis 6 ml 5 mm 4.6 mm 10 mm 1.6 mm 3.2 mm ÄKTA start* ÄKTAprime plus ÄKTA pure 25 ÄKTApurifier 10 plus ÄKTA pure 150 ÄKTApurifier 100 plus ÄKTA avant 25 ÄKTAexplorer 10 ÄKTA avant 150 ÄKTAexplorer 100 ÄKTAxpress ÄKTAmicro ÄKTApilot ÄKTA pilot 600 Recommended for system. Can be used with the system, but this combination might not give optimal results. * ÄKTA start is not specifically designed for HIC. However, HIC columns may be used in conditions where buffers are within specified system viscosity range and if gradient elution is not required. For optimized configuration for SEC columns, see cue card available from gelifesciences.com HiPrep 26 mm and HiLoad 26 mm SEC columns can be used with the system, but this combination is not optimal due to peak broadening (loss of resolution). Join ÄKTA club today gelifesciences.com/aktaclub KA SG 3

4 s for size exclusion chromatography, desalting/buffer exchange SEC allows separation of entities that differ in molecular size under mild conditions. The technique can be used for preparative protein purification and protein analysis, or for desalting and buffer exchange of samples. GE s SEC columns cover a molecular weight range from M r 100 to , from peptides to very large proteins, protein complexes, and viruses. Tip for choosing SEC columns Select the resin according to the fractionation range that covers the molecular weight values of interest in your sample. When your are interested in a specific component, select the resin where the target protein falls in the middle of the selectively curve. HiPrep Sephacryl, HiLoad Superdex, and Superdex 200 Increase columns Sample Fractionation range (M r 10 3 ) size, d 50V Sample > 0.5 ml: SEC columns for preparative purification HiLoad 16/600 Superdex 30 pg ml ml HiLoad 16 mm Superdex 30 prep grade HiLoad 26/600 Superdex 30 pg ml ml HiLoad 26 mm HiLoad 16/600 Superdex 75 pg ml ml HiLoad 16 mm Superdex 75 prep grade HiLoad 26/600 Superdex 75 pg ml ml HiLoad 26 mm HiLoad 16/600 Superdex 200 pg ml ml HiLoad 16 mm Superdex 200 prep grade HiLoad 26/600 Superdex 200 pg ml ml HiLoad 26 mm HiPrep 16/60 Sephacryl S 100 HR ml ml HiPrep 16 mm Sephacryl S 100 HR HiPrep 26/60 Sephacryl S 100 HR ml ml HiPrep 26 mm HiPrep 16/60 Sephacryl S 200 HR ml ml HiPrep 16 mm Sephacryl S 200 HR HiPrep 26/60 Sephacryl S 200 HR ml ml HiPrep 26 mm HiPrep 16/60 Sephacryl S 300 HR ml ml HiPrep 16 mm Sephacryl S 300 HR HiPrep 26/60 Sephacryl S 300 HR ml ml HiPrep 26 mm HiPrep 16/60 Sephacryl S 400 HR ml ml HiPrep 16 mm Sephacryl S 400 HR HiPrep 26/60 Sephacryl S 400 HR ml ml HiPrep 26 mm HiPrep 16/60 Sephacryl S 500 HR ml ml HiPrep 16 mm Sephacryl S 500 HR HiPrep 26/60 Sephacryl S 500 HR ml ml HiPrep 26 mm < µm µm µm µm µm µm µm (dextran) µm Median particle size of the cumulative distribution. 4 KA SG

5 Sample Fractionation range (M r 10 3 ) size, d 50V Sample < 0.5 ml: SEC columns for analysis or small scale preparative purification Superdex 30 Increase 3.2/ ml µl Precision 3.2 mm Superdex 30 Increase 10/300 GL 1 x 24 ml µl Tricorn 10 mm Superdex 75 Increase 3.2/ ml µl Precision 3.2 mm Superdex 75 Increase 5/150 GL 1 3 ml µl Tricorn 5 mm Superdex 75 Increase 10/300 GL 1 24 ml µl Tricorn 10 mm Superdex 200 Increase 3.2/ ml µl Precision 3.2 mm Superdex 200 Increase 5/150 GL 1 3 ml µl Tricorn 5 mm Superdex 200 Increase 10/300 GL 1 24 ml µl Tricorn 10 mm Superose 6 Increase 3.2/ ml µl Precision 3.2 mm Superose 6 Increase 5/150 GL 1 3 ml µl Tricorn 5 mm Superose 6 Increase 10/300 GL 1 24 ml µl Tricorn 10 mm Superose / ml µl Precision 3.2 mm Superose 12 10/300 GL 1 24 ml µl Tricorn 10 mm Superdex 30 Increase Superdex 75 Increase Superdex 200 Increase Superose 6 Increase to 12 9 µm to 12 9 µm to µm to µm Superose to µm Desalting/Buffer exchange columns HiPrep 26/10 Desalting 1 53 ml ml HiPrep 26 mm Sephadex 4 53 ml ml G 25 Fine HiTrap Desalting 1 5 ml 5 5 ml ml ml HiTrap 5 ml Sephadex G 25 Superfine ml ml 5 15 Median particle size of the cumulative distribution. Special pack size delivered on specific customer order. > 5 (exclusion limit) > 5 (exclusion limit) 2 to µm 2 to µm Superdex 200 Increase 10/300 GL column ( mm) in use on an ÄKTA pure system. KA SG 5

6 s for ion exchange chromatography IEX separates proteins with differences in net surface charge. The technique is commonly used for the purification of untagged or native proteins. IEX is also an excellent technique for purity and heterogeneity analysis. Tips for choosing IEX chromatography columns Begin with a strong ion exchanger, allowing work over a broad range during the initial method development. Consider using a weak exchanger if the selectivity of the strong ion exchanger is unsatisfactory. Use one of the HiTrap selection kits to screen for the most suitable ion exchanger. Keep in mind that smaller bead size delivers increased resolution but higher back pressure. HiTrap, RESOURCE, and Tricorn IEX columns s per pack bed Typical loading range size, d 50V Strong anion exchangers Mono Q PC 1.6/ ml < Precision 1.6 mm Mono Q 2 to µm Mono Q 5/50 GL 1 1 ml Tricorn 5 mm Mono Q 4.6/100 PE ml Tricorn 4.6 mm Mono Q 10/100 GL 1 8 ml Tricorn 10 mm Mono Q HR 16/ ml HR 16 mm SOURCE 15Q 4.6/100 PE ml Tricorn 4.6 mm SOURCE 15Q 2 to µm RESOURCE Q, 1 ml 1 1 ml RESOURCE 1 ml RESOURCE Q, 6 ml 1 6 ml RESOURCE 6 ml HiTrap Q HP 1 1 ml 5 1 ml (HSA) 1 4 HiTrap 1 ml Q Sepharose High 5 5 ml (HSA) HiScreen Q HP ml (HSA) HiScreen 4.7 ml HiPrep Q HP 16/ ml (HSA) 3 5 HiPrep 16 mm HiTrap Capto Q ImpRes 5 1 ml (BSA) 1 4 HiTrap 1 ml Capto Q ImpRes 5 5 ml (BSA) HiScreen Capto Q ImpRes ml (BSA) HiScreen 4.7 ml HiPrep Q FF 16/ ml (HSA) 5 10 HiPrep 16 mm Q Sepharose HiTrap Q FF 5 1 ml (HSA) 1 4 HiTrap 1 ml 5 5 ml (HSA) HiScreen Q FF ml (HSA) HiScreen 4.7 ml 2 to µm 2 to µm 2 to µm HiPrep Q XL 16/ ml (BSA) 5 10 HiPrep 16 mm Q Sepharose XL 2 to µm HiTrap Q XL 5 1 ml (BSA) 1 4 HiTrap 1 ml 5 5 ml (BSA) HiTrap Capto Q 5 1 ml (BSA) 1 4 HiTrap 1 ml Capto Q 2 to µm 5 5 ml (BSA) HiScreen Capto Q ml (BSA) HiScreen 4.7 ml Median particle size of the cumulative distribution. This column format is compatible with ÄKTAexplorer, ÄKTApurifier, ÄKTA pure, and ÄKTA avant. 6 KA SG

7 Typical loading range size, d 50V Weak anion exchangers HiPrep DEAE FF 16/ ml (HSA) 5 10 HiPrep 16 mm DEAE 2 to µm HiTrap DEAE FF 5 1 ml (HSA) 1 4 HiTrap 1 ml Sepharose 5 5 ml (HSA) HiScreen DEAE FF ml (HSA) HiScreen 4.7 ml HiTrap ANX FF (high sub) 5 1 ml (BSA) 1 4 HiTrap 1 ml ANX Sepharose 3 to µm 5 5 ml (BSA) HiTrap Capto DEAE 5 1 ml (ovalbumin) 1 4 HiTrap 1 ml Capto DEAE 2 to µm 5 5 ml (ovalbumin) HiScreen Capto DEAE ml (ovalbumin) HiScreen 4.7 ml Strong cation exchangers Mono S PC 1.6/ ml < Precision 1.6 mm Mono S 2 to µm Mono S 5/50 GL 1 1 ml Tricorn 5 mm Mono S 4.6/100 PE ml Tricorn 4.6 mm Mono S 10/100 GL 1 8 ml Tricorn 10 mm Mono S HR 16/ ml HR 16 mm SOURCE 15S 4.6/100 PE ml Tricorn 4.6 mm SOURCE 15S 2 to µm RESOURCE S, 1 ml 1 1 ml RESOURCE 1 ml RESOURCE S, 6 ml 1 6 ml RESOURCE 6 ml HiPrep SP HP 16/ ml HiPrep 16 mm SP Sepharose 4 to µm HiTrap SP HP 1 1 ml (ribonuclease A) 150 (ribonuclease A) 1 4 HiTrap 1 ml High 5 1 ml ml (ribonuclease A) HiScreen SP HP ml (ribonulease A) HiScreen 4.7 ml HiTrap Capto SP ImpRes 5 1 ml (lysozyme) 1 4 HiTrap 1 ml Capto SP 4 to µm 5 5 ml (lysozyme) ImpRes HiScreen Capto SP ImpRes ml (lysozyme) HiScreen 4.7 ml HiTrap Capto S ImpAct 5 1 ml (antibodies) 1 4 HiTrap 1 ml Capto S ImpAct 4 to µm 5 5 ml (antibodies) HiScreen Capto S ImpAct ml (antibodies) HiScreen 4.7 ml HiPrep SP FF 16/ ml (ribonuclease A) 5 10 HiPrep 16 mm SP Sepharose 4 to µm HiTrap SP FF 5 1 ml (ribonuclease A) 1 4 HiTrap 1 ml 5 5 ml (ribonuclease A) HiScreen SP FF ml (ribonulease A) HiScreen 4.7 ml HiPrep SP XL 16/ ml (lysozyme) 5 10 HiPrep 16 mm SP Sepharose 4 to µm HiTrap SP XL 5 1 ml (lysozyme) 1 4 HiTrap 1 ml XL 5 5 ml (lysozyme) HiTrap Capto S 5 1 ml (lysozyme) 1 4 HiTrap 1 ml Capto S 4 to µm 5 5 ml (lysozyme) HiScreen Capto S ml (lysozyme) HiScreen 4.7 ml Weak cation exchangers HiPrep CM FF 16/10 HiTrap CM FF 1 20 ml 5 1 ml (ribonuclease A) 150 (ribonuclease A) HiPrep 16 mm HiTrap 1 ml CM Sepharose 5 5 ml (ribonuclease A) 6 to µm Ion exchange selection kits HiTrap IEX Selection Kit 7 1 ml depends on resin 1 4 HiTrap 1 ml Various FF and XL resins HiTrap Capto IEX Selection Kit 5 1 ml depends on resin 1 4 HiTrap 1 ml Various Capto resins depends on resin depends on resin 34 or 90 µm 75 or 90 µm Median particle size of the cumulative distribution. This column format is compatible with ÄKTAexplorer, ÄKTApurifier, ÄKTA pure and ÄKTA avant. SP Sepharose, SP Sepharose XL, CM Sepharose, Q Sepharose, Q Sepharose XL, DEAE Sepharose, ANX Sepharose 4. Capto Q, Capto S, Capto DEAE, Capto MMC, Capto adhere. KA SG 7

8 s for purification Tagged protein typically requires the use of affinity chromatography (AC) in the first purification step. AC separates proteins on the basis of a reversible interaction between the target protein (or group of proteins) and a specific ligand attached to a chromatography matrix. Tips for choosing histidine tagged (his tagged) protein purification columns When high binding capacity is the priority: choose Ni Sepharose products. (HP as a first choice, FF for batch/gravity or scale up). When nickel leakage is an issue: Choose Ni Sepharose excel products. When high purity is the priority: Choose TALON Superflow products. HiTrap TALON crude, HisTrap FF crude, and HisTrap excel columns Typical loading range size, d 50V His purification columns HisTrap HP 1 1 ml ml (at least) (his) ml ml ml (at least) (his) ml HisTrap FF 5 1 ml (his) ml ml (his) ml HisTrap FF crude 1 1 ml (his) ml ml ml (his) ml HisPrep FF 16/ ml (his) 6 HiScreen Ni FF ml mg (his) 6 HisTrap excel 1 1 ml (his) ml ml (his) 6 HiTrap TALON crude 1 1 ml (his) ml ml (his) 6 Median particle size of the cumulative distribution. Special pack size delivered on specific customer order. 1 4 HiTrap 1 ml Ni Sepharose High 1 4 HiTrap 1 ml Ni Sepharose HiTrap 1 ml 5 10 HiPrep 16 mm HiScreen 4.7 ml 1 4 HiTrap 1 ml Ni Sepharose excel 1 4 HiTrap 1 ml TALON Superflow 3 to µm 3 to µm 2 to µm 3 to µm 8 KA SG

9 Typical loading range size, d 50V GST purification columns GSTrap HP 5 1 ml (recombinant GST) HiTrap 1 ml Glutathione Sepharose 1 5 ml HiTrap 5 ml High (recombinant GST) 5 5 ml GSTrap FF 2 1 ml ml (recombinant GST) 1 5 ml ml (recombinant GST) GSTPrep FF 16/ ml (recombinant GST) GSTrap 4B 1 1 ml ml (recombinant GST) 1 5 ml ml (recombinant GST) HiTrap 1 ml Glutathione Sepharose HiTrap 5 ml 2 10 HiPrep 16 mm HiTrap 1 ml Glutathone Sepharose 4B HiTrap 5 ml 3 to µm 3 to µm 4 to µm Strep tag II purification columns StrepTrap HP 1 1 ml 5 1 ml (Strep tag II protein) 1 5 ml (Strep 5 5 ml tag II protein) MBP purification columns MBPTrap HP 1 1 ml (MBP 5 1 ml ) 1 5 ml (MBP 5 5 ml ) HiTrap 1 ml HiTrap 5 ml StrepTactin Sepharose High HiTrap 1 ml HiTrap 5 ml Dextrin Sepharose High > 7 34 µm > 7 34 µm Immobilized metal ion affinity (IMAC) columns to be charged with the metal of your choice HiTrap IMAC HP 5 1 ml (his) 6 tagged protein (Ni + charged) 5 5 ml (his) 6 tagged protein (Ni + charged) HiTrap IMAC FF 5 1 ml (his) 6 tagged protein (Ni + charged) 5 5 ml (his) 6 tagged protein (Ni + charged) HiPrep IMAC FF 16/ ml (his) 6 tagged protein (Ni + charged) HiScreen IMAC FF ml (his) 6 tagged protein (Ni + charged) HiTrap Chelating HP 5 1 ml (his) 6 tagged protein (Ni + charged) 1 5 ml (his) 6 tagged 5 5 ml protein (Ni + charged) ml HiTrap 1 ml IMAC Sepharose High 1 4 HiTrap 1 ml IMAC Sepharose HiPrep 16 mm HiScreen 4.7 ml 1 4 HiTrap 1 ml Chelating Sepharose High 3 to µm 3 to µm 3 to µm Median particle size of the cumulative distribution. Special pack size delivered on specific customer order. KA SG 9

10 s for antibody purification Polyclonal antibodies, monoclonal antibodies (mab), and antibody fragments are usually purified by affinity chromatography. s containing an immobilized ligand (e.g., protein A, protein G, or protein L) are used to capture antibodies and antibody fragments. Tips for choosing protein A antibody purification columns Choose Protein A Sepharose High products as a first choice for routine purification. Choose MabSelect PrismA if you need to purify different types of antibodies or if you need to scale up. HiTrap Protein A HP columns, 1 ml and 5 ml Typical loading range size, d 50V Protein A for purifying human IgG HiTrap Protein A HP 1 1 ml (human IgG) HiTrap 1 ml Protein A Sepharose 3 to µm 2 1 ml High 5 1 ml ml (human IgG) ml HiTrap rprotein A FF 2 1 ml (human IgG) HiTrap 1 ml rprotein A Sepharose 3 to µm 5 1 ml ml (human IgG) ml HiTrap MabSelect 5 1 ml (human IgG) HiTrap 1 ml MabSelect 3 to µm 1 5 ml (human IgG) ml HiScreen MabSelect ml human IgG) HiScreen 4.7 ml HiTrap MabSelect Xtra 5 1 ml (human IgG) HiTrap 1 ml MabSelect Xtra 3 to µm 1 5 ml (human IgG) ml HiScreen MabSelect Xtra ml (human IgG) HiScreen 4.7 ml HiTrap MabSelect PrismA 1 1 ml (human IgG) HiTrap 1 ml MabSelect PrismA 3 to 12 ~60 µm 5 1 ml ml (human IgG) ml HiScreen ml (human IgG) HiScreen 4.7 ml MabSelect PrismA HiTrap MabSelect SuRe 1 1 ml (human IgG) HiTrap 1 ml MabSelect SuRe 3 to µm 5 1 ml ml (human IgG) ml HiScreen MabSelect SuRe ml (human IgG) HiScreen 4.7 ml HiTrap 5 1 ml (human IgG) HiTrap 1 ml MabSelect SuRe LX 3 to µm MabSelect SuRe LX 5 5 ml (human IgG) 2. HiScreen MabSelect ml (human IgG) HiScreen 4.7 ml SuRe LX HiTrap MabSelect 5 1 ml (human IgG) HiTrap 1 ml MabSelect SuRe pcc 3 to µm SuRe pcc 1 5 ml (human IgG) 2. Median particle size of the cumulative distribution. This product is part of our Custom Design Media program. Delivery time may be longer than for standard products. 10 KA SG

11 Typical loading range size, d 50V Protein G for purifying IgG from most species, including rat HiTrap Protein G HP 1 1 ml 2 1 ml (human IgG) 5 1 ml ml ml (human IgG) 1 4 HiTrap 1 ml Protein G Sepharose High 3 to 9 34 µm Purification of antibody fragments HiTrap Protein L 1 1 ml (Fab) 1 4 HiTrap 1 ml Capto L 2 to µm 5 1 ml ml (Fab) 5 5 ml HiScreen Capto L ml (Fab) HiScreen 4.7 ml Purification of IgM and IgY HiTrap IgM Purification HP 1 1 ml (human IgM) 1 4 HiTrap 1 ml 2 mercaptopyridine Sepharose High Perfomance (2 mg/ml) HiTrap IgY Purification HP 1 5 ml (IgY) 2 mercaptopyridine Sepharose High (3.5 mg/ml) 3 to µm Affinity columns for specific groups of biomolecules HiTrap Blue HP columns Typical loading range size, d 50V Group specific affinity columns HiTrap Heparin HP 5 1 ml (bovine 1 4 HiTrap 1 ml Heparin Sepharose 5 to µm antithrombin III) High 1 5 ml (bovine 5 5 ml antithrombin III) HiPrep Heparin FF 16/ ml (bovine 5 10 HiPrep 16 mm Heparin Sepharose 4 to µm antithrombin III) HiTrap Blue HP 5 1 ml (HSA) 1 4 HiTrap 1 ml Blue Sepharose 4 to µm 1 5 ml (HSA) High HiScreen Capto Blue ml HSA HiScreen 4.7 ml Capto Blue 3 to µm HiScreen Blue FF ml (HSA) HiScreen 4.7 ml Blue Sepharose 3 to µm HiTrap Con A 4B 5 1 ml (porcine 1 4 HiTrap 1 ml Con A Sepharose 4 to 9 90 µm thyroglobulin) 4 B 5 5 ml (porcine thyroglobulin) HiTrap Streptavidin HP 5 1 ml > 300 nmol biotin 1 4 HiTrap 1 ml Strepatvidin Sepharose High 4 to 9 34 µm HiTrap Benzamidine FF (high sub) 2 1 ml (trypsin) 1 4 HiTrap 1 ml Benzamidine 5 1 ml Sepharose (high 1 5 ml (trypsin) sub) Preactivated columns for preparing your own affinity column HiTrap NHS activated HP 5 1 ml application dependent 5 5 ml application dependent 1 4 HiTrap 1 ml NHS activated Sepharose High 2 to 8 90 µm Ligand dependent Median particle size of the cumulative distribution. 34 µm KA SG 11

12 s for hydrophobic interaction chromatography HIC separates proteins according to differences in the surface hydrophobicities by utilizing a reversable interaction between the proteins and the hydrophobic ligand of a HIC resin. Tips for choosing HIC chromatography columns The hydrophobicity of a protein is difficult to determine. Screen for the most suitable resin for each application using the HiTrap HIC Selection Kit or RESOURCE HIC Test kit. Otherwise, a good practice is to start with a phenyl ligand. Select the resin that gives the optimal resolution and loading capacity at a low salt concentration. Hydrophobic ligands that vary according to their degree of hydrophobicity are available: Increasing hydrophobicity : butyl S ether isopropyl butyl octyl phenyl. HiTrap HIC Selection Kit Ligand density size, d 50V HiTrap Butyl HP 5 1 ml µmol/ml 1 4 HiTrap 1 ml Butyl Sepharose 3 to µm 5 5 ml High HiScreen Butyl HP ml HiScreen 4.7 ml HiTrap Butyl S FF 5 1 ml µmol/ml 1 4 HiTrap 1 ml Butyl S Sepharose 3 to µm HiScreen Butyl S FF ml HiScreen 4.7 ml 6 HiTrap Butyl FF 5 1 ml µmol/ml 1 4 HiTrap 1 ml Butyl Sepharose 4 3 to µm 5 5 ml HiPrep Butyl FF 16/ ml HiPrep 16 mm HiScreen Butyl FF ml HiScreen 4.7 ml HiScreen Capto Butyl ml µmol/ml HiScreen 4.7 ml Capto Butyl 3 to µm HiTrap Phenyl HP 5 1 ml µmol/ml 1 4 HiTrap 1 ml Phenyl Sepharoe 3 to µm 5 5 ml High HiPrep Phenyl HP 16/ ml HiPrep 16 mm HiScreen Phenyl HP ml HiScreen 4.7 ml HiTrap Phenyl FF (low sub) 5 1 ml µmol/ml 1 4 HiTrap 1 ml Phenyl Sepharose 3 to µm 5 5 ml (low sub) HiScreen Phenyl FF ml HiScreen 4.7 ml (low sub) HiTrap Phenyl FF (high sub) 5 1 ml µmol/ml 1 4 HiTrap 1 ml Phenyl Sepharose 3 to µm 5 5 ml (high sub) HiPrep Phenyl FF (high sub) 1 20 ml HiPrep 16 mm 16/10 HiScreen Phenyl FF ml HiScreen 4.7 ml (high sub) HiScreen Capto Phenyl ml µmol/ml HiScreen 4.7 ml Capto Phenyl (high 3 to µm (high sub) sub) HiTrap Octyl FF 5 1 ml µmol/ml 1 4 HiTrap 1 ml Octyl Sepharose 4 3 to µm 5 5 ml HiPrep Octyl FF 16/ ml HiPrep 16 mm HiScreen Octyl FF ml HiScreen 4.7 ml SOURCE 15 PHE 4.6/100 PE ml Not available 1 5 Tricorn 4.6 mm SOURCE 15 PHE 2 to µm RESOURCE ETH 1 1 ml Not available RESOURCE 1 ml SOURCE ETH 2 to µm RESOURCE ISO 1 1 ml Not available RESOURCE 1 ml SOURCE ISO 2 to µm RESOURCE PHE 1 1 ml Not available RESOURCE 1 ml SOURCE PHE 2 to µm 12 KA SG

13 Ligand density size, d 50V HIC selection kits HiTrap HIC Selection Kit 7 1 ml depends on resin 1 4 HiTrap 1 ml Various 3 to or 34 µm RESOURCE HIC Test Kit 3 1 ml Not available RESOURCE 1 ml Various 2 to µm Median particle size of the cumulative distribution. HiTrap HIC Selection Kit content: Seven HITrap columns packed with the following resins: Phenyl Sepharose High, Butyl Sepharose High, Phenyl Sepharose 6 (low sub), Phenyl Sepharose 6 (high sub), Butyl S Sepharose 6, Butyl Sepharose 4, and Octyl Sepharose 4. RESOURCE HIC Test Kit content: RESOURCE PHE, RESOURCE ISO, and RESOURCE ETH. s for multimodal chromatography Typical loading range HiTrap Capto 5 1 ml mab 1 4 HiTrap 1 ml Capto adhere ImpRes adhere HiScreen Capto ml mab HiScreen 4.7 ml ImpRes adhere ImpRes HiTrap Capto adhere 5 1 ml Not available 1 4 HiTrap 1 ml Capto 5 5 ml Not available adhere HiScreen Capto adhere ml Not available HiScreen 4.7 ml HiTrap Capto MMC ImpRes 5 1 ml mab 1 4 HiTrap 1 ml Capto MMC HiScreen Capto ml mab HiScreen 4.7 ml ImpRes MMC ImpRes HiTrap Capto MMC 5 1 ml (BSA at 30 ms/cm) 5 5 ml (BSA at 30 ms/cm) HiScreen Capto MMC ml (BSA at 30 ms/cm) 1 4 HiTrap 1 ml Capto MMC HiScreen 4.7 ml HiScreen Capto MMC and HiTrap Capto MMC 1 ml and 5 ml HiTrap Capto Core ml ovalbumin HiTrap 5 ml Capto HiScreen Capto Core ml ovalbumin HiScreen 4.7 ml Core 700 size, d 50V 3 to µm 3 to µm 3 to µm 2 to µm 3 to µm s for reversed phase chromatography Typical loading range SOURCE 15RPC ST 4.6/ ml (BSA) 2 5 Stainless steel column RESOURCE RPC, 1 ml 1 1 ml (BSA) 2 10 RESOURCE 1 ml RESOURCE RPC, 3 ml 1 3 ml (BSA) 2 10 RESOURCE 3 ml Median particle size of the cumulative distribution. This column format is compatible with ÄKTAexplorer, ÄKTAmicro, ÄKTApurifier, ÄKTA pure, and ÄKTA avant. SOURCE 15RPC SOURCE15 RPC ST 4.6/100 columns size, d 50V 1 to µm KA SG 13

14 Check out GE s complete range of ÄKTA systems for research and process development With ÄKTA protein purification systems, backed by GE s columns and chromatography resins, you are taken reliably from lab bench to pilot and production scale. Way of working ÄKTA start ÄKTAprime plus ÄKTAxpress ÄKTA pure ÄKTA avant Simple, one step desalting, buffer exchange Automated and reproducible protein purification using all common techniques including support for gradient elution Research Software compatible with regulatory requirements, e.g., GLP Method development and optimization using design of experiments (DoE) Automatic buffer preparation including scouting Automatic resin or column scouting Automatic multistep purification Scale up, process development () * * Max. operating pressure (MPa) * 5 20* 5 Software for system control and data handling UNICORN start PrimeView UNICORN 5 UNICORN 6 or later UNICORN 6 or later included optional * ÄKTA pure 25 and ÄKTA avant 25 ÄKTA pure 150 and ÄKTA avant 150 A specific software version might be needed for the chosen system. See the web page for each respective system at gelifesciences.com/aktalabsystems With PrimeView, you can monitor results and evaluate data but not create methods nor control the system. GLP = good laboratory practice 1 MPa = 10 bar, 145 psi Process development Upgrade your ÄKTA system Since the 1990 s, ÄKTA systems have offered versatile and reliable protein purification. As a consequence of the renewal of the ÄKTA system platform, production of ÄKTAexplorer, ÄKTApurifier, ÄKTAfplc, and ÄKTAmicro has been stopped. To improve your protein purification output, we recommend upgrading to ÄKTA avant and ÄKTA pure, as outlined in this table. Discontinued system Upgrade to ÄKTAexplorer 10 ÄKTAexplorer 10S ÄKTAexplorer 100 ÄKTAexplorer 100 Air ÄKTAfplc ÄKTApurifier UPC 10 ÄKTApurifier 10 ÄKTApurifier 10 plus ÄKTA avant ÄKTA avant ÄKTA pure 25 L ÄKTA pure 25 L1 ÄKTA pure 25 M ÄKTA pure 25 M1 ÄKTA pure 25 M ÄKTApurifier UPC 100 ÄKTA pure 150 L ÄKTApurifier 100 ÄKTApurifier 100 plus ÄKTAmicro ÄKTA pure 150 M ÄKTA pure 150 M3 ÄKTA pure 25 with microgram scale purification flow path * * Contact your local sales representative for further information. 14 KA SG

15 Get further guidance on product selection Purify App column and resin interactive selection tool Download the app at gelifesciences.com/purify ÄKTA system accessories app Download the app at gelifesciences.com/aktaapps Download GE s protein purification handbooks Protein purification handbooks In depth theory and practical tips to help make your experiments a success Available from gelifesciences.com/proteinhandbooks KA SG 15

16 gelifesciences.com/proteinresearch GE, the GE Monogram, ÄKTA, ÄKTApilot, Capto, GSTrap, GSTPrep, HiLoad, HiTrap, HiPrep, HiScreen, HisPrep, HisTrap, MabSelect, MabSelect Xtra, MabSelect SuRe, MBPTrap, Mono Q, Mono S, PrimeView, RESOURCE, Sephacryl, Sephadex, Sepharose, SOURCE, StrepTrap, Superdex, Superose, UNICORN, and Tricorn are trademarks of General Electric Company. TALON is a trademark of Clontech Laboratories, Inc. Strep Tactin and Strep tag are trademarks of IBA GmbH. Superflow is a trademark of Sterogene Biospearations Inc. All other trademarks are the property of their respective owners. IMAC Sepharose products, Ni Sepharose products and Fe Sepharose products: These products are sold under a license from Sigma Aldrich under patent number EP (Metal chelating compositions) and equivalent patents and patent applications in other countries. Protein L: Protein L is covered by US 6, 822,075, US 6,162, 903, US 6,884,629 and equivalent patents and patent applications in other countries owned by Affitech AS and exclusively licensed to GE Healthcare. StrepTrap HP and StrepTactin Sepharose High : These products are covered by US patent number 6,103,493 and equivalent patents and patent applications in other countries. The purchase of StrepTrap HP and StrepTactin Sepharose High includes a license under such patents for non profit and in house research only. Please contact IBA (info@iba go.com) for further information on licenses for commercial use of StrepTactin General Electric Company. All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information. GE Healthcare Bio Sciences AB, Björkgatan 30, Uppsala, Sweden GE Healthcare Bio Sciences Corp., 100 Results Way, Marlborough, MA 01752, USA GE Healthcare Europe GmbH, Munzinger Strasse 5, D Freiburg, Germany GE Healthcare Japan Corp., Sanken Bldg., , Hyakunincho Shinjuku ku, Tokyo , Japan GE Healthcare UK Limited, Amersham Place, Little Chalfont, Buckinghamshire, HP7 9NA, UK For local office contact information, visit gelifesciences.com/contact AL KA SG