GE Healthcare. ÄKTAxpress protein purification that keeps pace with your research

Transcription

1 GE Healthcare ÄKTAxpress protein purification that keeps pace with your research

2 Keeping pace with today s purification challenges ÄKTAxpress TM is a complete, intelligent, and robust solution for multidimensional purification of affinity-tagged proteins and antibodies. Automated multistep purification provides the highest possible protein purity. ÄKTAxpress can run multiple samples providing both the required purity and high throughput. Robotic systems are limited to a single chromatography step and sacrifice purity to achieve high throughput. Now it is possible to purify proteins expressed at all levels, even proteins expressed at low levels can be purified. Dual-tagged proteins are purified easily and you have the option of on- or offcolumn tag cleavage. 2

3 Important benefits Importance to your research Throughput Ability to automatically run up to 8 samples per TWIN, up to 48 samples with 12 modules, with little hands-on time needed. Maximize productivity, system can grow with increasing throughput needs. Capacity From 1 mg to 50+ mg yields of pure protein. Purity Automated multistep purification provides the highest possible target protein purity >95%. Flexibility Intelligence UNICORN wizard Reliability Different protocols can be run simultaneously on different modules. Hardware, software and media designed to work together eliminates guesswork. No chromatography expertise is required. Intelligent software user can program the system by ticking off boxes in the wizard, no purification expertise required, complete purification documentation and evaluation. Industry standard for protein purification, with over installed systems well established service and support experience. 3

4 ÄKTAxpress TWIN module 4

5 Intelligence Hardware, software and media designed to work together. High throughput Ability to automatically run up to eight samples per TWIN, with little hands-on time needed. High capacity From 1 mg to 50+ mg yields of pure protein. Purity Automated multistep purification provides the highest possible target protein purity >95%. Flexibility Different protocols can be run simultaneously on different modules. Reliability Industry standard for protein purification, with over installed systems well estab lished service and support experience. 5

6 A complete solution UNICORN TM software allows easy system set-up and unattended protein purification, with full documentation and evaluation. Chromatography expertise is not required. An extensive range of prepacked columns for purification of tagged proteins and monoclonal antibodies (MAbs) are designed to work together with the system. The method wizard makes it easy to create a method plan for a purification protocol. It guides you with checkboxes and pull down menus, making it simple to create protocols with up to four different purification steps. Simply decide the number of purification steps and choose the appropriate columns from the column list. There are preprogrammed default values for all running parameters. An advanced zone in the method wizard gives you the option to change the default settings. protocols can be run even though a single computer controls them. Each module can purify up to four different proteins serially in one run. If 12 modules are run in parallel, up to 48 samples can be purified in one run. New algorithms in the embedded software allow the detection and storage of intermediate peaks in capillary loops, allowing the system to automatically wash, equilibrate, elute, or re-inject samples on the different columns connected to the system. Each module has a fixed configuration that consists of the system pump, an air sensor, a UV detector (280 or 254 nm), a conductivity detector and an integrated fraction collector for 24 or 96 deep-well plates. All necessary items, such as valves, sample loops and tubing, are pre-installed. An optional flask holder is also available. Robust algorithms and combinations of watch commands make it possible to automatically process interesting protein peaks at different stages in the purification process. ÄKTAxpress has a modular design that allows for a high degree of adaptability. A TWIN pack can be upgraded with separate modules, and up to 12 modules can be controlled from one computer. Each module is run independently, and different 6

7 ÄKTAxpress 12 modules High throughput Ability to automatically run up to 48 samples with 12 modules, with little hands-on time needed.

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10 Tagged protein purification highest possible purity for structural studies ÄKTAxpress can handle protocols with up to four purification steps: affinity (AC), desalting (DS), ion exchange (IEX), gel filtration (GF) and optional tag removal. Dual-tagged proteins can be purified using protocols that include two AC steps. Flexible peak selection enables successful purification of proteins expressed at low levels. Some of the supported protocols for tagged protein purification Protocol Typical run time Number of samples/module AC 2.5 hours 4 AC/IEX-DS 4.5 hours 4 AC/IEX-GF 10.5 hours 4 AC/IEX-DS-AC/IEX 6 hours 3 AC/IEX-DS-AC/IEX-DS 8.5 hours 3 AC/IEX-DS-AC/IEX-GF 8 hours 2 DS 45 min. 4 GF 6 hours 4 Automated tag removal An optional tag removal step can be integrated in the on-column purification protocol. The method works for a broad range of proteases, both untagged and tagged. There is also an option for off-column tag removal. Protease injection Cleaved protein elution Sample loading Incubation Regeneration Collection in loop 10 Further purification

11 Save time and effort The system is designed to eliminate the manual steps involved in multistep purifications. Traditional methods With ÄKTAxpress When working with tagged proteins Sample preparation Running multistep protocols Harvest your cells, sonicate, clarify sample, purify. Pool fractions between each step, start next purification step and apply sample to the column. Using HisTrap FF crude apply unclarified sample directly on column. Save up to 1 hour/sample. Automatic sample loading of collected peak onto next column. Save up to 12 hours when running a four-step protocol. Skip a step in sample prep When using the HisTrap FF crude column, you can apply unclarified cell extract directly onto the column and save min. per sample without risking any clogging of the column. Without HisTrap FF crude Cell culture > Sonication > Sample prep > Load sample > Purified protein With HisTrap FF crude Cell culture > Sonication > Load sample > Purified protein 11

12 ÄKTAxpress results for tagged protein purification Four-step purification (AC-DS-IEX-GF) of APB7, a protein of 28 kda AC: DS: IEX: GF: HisTrap HP, 5 ml HiPrep 26/10 Desalting 2 x HiTrap Q HP, 1 ml HiLoad 16/60 Superdex 75 pg Four-step purification with tag removal included AC: DS: IEX: GF: HisTrap HP, 5 ml HiPrep 26/10 Desalting RESOURCE Q, 6 ml HiLoad 16/60 Superdex 75 PG Protease: AcTEV histidine-tagged Lane 1 Low molecular weight marker 2 Start sample 3 Flowthrough 4 Purified, cleaved APC 1040 (M r ) after AC-DS-IEX-GF 5 Reference: uncleaved APC 1040 (M r ) 12

13 Dual-tagged proteins enable more targeted affinity purification of full length proteins, if the tags are at different terminals of the protein. Protocols containing two affinity steps have been developed to support purification of dualtagged proteins. mau AC DS AC DS AC: DS: AC: DS: HisTrap HP, 5ml HiPrep 26/10 Desalting GSTrap 4B, 1ml HiPrep 26/10 Desalting ml Lane 1 Low molecular weight marker 2 Start sample 3 Flowthrough after AC 4 Green fluorescent protein tagged with gluthathione S-transferase at the N-terminal and with (histidine) 6 at the C-terminal (GST- GFP-His) (M r ~51 000) 5 Flowthrough after AC 6 Purified His-GFP-GST after AC 7 Flowthrough after AC 8 Purified His-GFP-GST after AC 9 Low molecular weight marker

14 MAb purification highest possible purity for preclinical screening Supported protocols for MAb purification Protocol No. of samples/module Affinity chromatography 4 Affinity chromatography followed by desalting, one single desalting column Affinity chromatography with individual desalting columns for each sample Affinity chromatography followed by gel filtration, one single gel filtration column Affinity chromatography with individual gel filtration columns for each sample Note: Always use a separate affinity column for each sample. Alkali stable HiTrap MabSelect SuRe The same cleaning procedure using 0.5 M NaOH is applicable for both the system and column without loss of binding capacity. Automated neutralization Choosing the desalting step automatically places the MAb in the appropriate buffer, avoiding degradation of the monoclonal antibody. 14

15 ÄKTAxpress results for MAb Two-step AC-DS of human IgG 1 from hybridoma cell culture supernatant. AC: HiTrap MabSelect SuRe 5 ml DS: HiPrep 26/10 Desalting mau AC DS ml Automated cleaning procedure between samples Extended washing procedures enable automatic processing of a large number of samples and minimize the risk of cross contamination. 15

16 One point of control The ÄKTAxpress system is controlled by UNICORN version 5.11 or higher. Algorithms and complex combinations of watch commands have been developed and optimized to enable intelligent automation and robust peak handling functions. Now even the most difficult of proteins can be tackled with ÄKTAxpress. The method wizard guides you through each stage of the method programming process. A method plan is generated, based on the answers to a series of questions. Columns are selected from the column library which includes all the prepacked columns designed to work with the intended application. Optimized values for all running parameters are used as a default to make it easy to create new method plans. An advanced zone within the wizard makes it possible to change default settings in order to further optimize the purification of your samples. 1 protocol Easy to create a method When you enter the main menu of the wizard, you can choose to make a method plan for a purification and/or different maintenance activities. 2 list. Pick what you want when it s ready To prepare a purification method plan, all you need to do is select what purification protocol you want to run and choose the appropriate columns from the column The rest is up to the wizard. 16

17 3 More advanced selection You can adjust running parameters in the advanced zone of the wizard if needed. 5 module Data in real time; monitor what s happening on all modules You get data in real time while you run the method, and you can follow what is happening in detail on one or have an overall status of all running modules. 4 Bar code data entry When starting a run, you assign the method plan to your different system modules and enter the number of samples you want to run per module. Sample data can be entered either manually or by using a bar code reader. 6 optimized Evaluation and creating reports Results are displayed in a way that makes evaluation simple. Reports are easily created by using the formats. 17

18 Ordering information ÄKTAxpress Related products (cont.) ÄKTAxpress TWIN system ÄKTAxpress module for system extension ÄKTAxpress software (includes upgrade & MAb) Related products Gel filtration HiLoad TM 16/60 Superdex TM 75 pg, 1 x 120 ml HiLoad 16/60 Superdex 200 pg, 1 x 120 ml HiLoad 26/60 Superdex 75 pg, 1 x 320 ml HiLoad 26/60 Superdex 200 pg, 1 x 320 ml HiPrep TM 16/60 Sephacryl S-100 HR, 1 x 120 ml HiPrep 16/60 Sephacryl S-200 HR, 1 x 120 ml HiPrep 16/60 Sephacryl S-300 HR, 1 x 120 ml HiPrep 26/60 Sephacryl S-100 HR, 1 x 320 ml HiPrep 26/60 Sephacryl S-200 HR, 1 x 320 ml HiPrep 26/60 Sephacryl S-300 HR, 1 x 320 ml Desalting HiPrep 26/10 Desalting, 1 x 53 ml HiPrep 26/10 Desalting, 4 x 53 ml HiTrap TM Desalting, 5 x 5 ml Affinity chromatography HisTrap HP, 5 x 1 ml HisTrap HP, 5 x 5 ml HiTrap MabSelect SuRe TM 5 x 1 ml HiTrap MabSelect SuRe 1 x 5ml HiTrap MabSelect SuRe 5 x 5 ml HiTrap Chelating HP, 5 x 1 ml HiTrap Chelating HP, 5 x 5 ml GSTrap TM HP, 5 x 1 ml GSTrap HP, 5 x 5 ml GSTrap FF, 5 x 1 ml GSTrap FF, 5 x 5 ml GSTrap 4B, 5 x 1 ml GSTrap 4B, 5 x 5 ml Ion exchange chromatography HiTrap SP HP, 5 x 1 ml HiTrap SP HP, 5 x 5 ml HiTrap Q HP, 5 x 1 ml HiTrap Q HP, 5 x 5 ml RESOURCE TM Q, 1 x 1 ml RESOURCE Q, 1 x 6 ml RESOURCE S, 1 x 1 ml RESOURCE S, 1 x 6 ml Mono Q TM 5/50 GL, 1 x 1 ml Mono S TM 5/50 GL, 1 x 1 ml

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20 GE Healthcare Bio-Sciences AB Björkgatan 30 SE Uppsala Sweden All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. GE Healthcare reserves the right, subject to any regulatory and contractual approval, if required, to make changes in specifications and features shown herein, or discontinue the product described at any time without notice or obligation. Contact your local GE Healthcare representative for the most current information General Electric Company All rights reserved. GE Healthcare Bio-Sciences AB, a General Electric Company. GE Healthcare Europe GmbH Munzinger Strasse 5 D Freiburg, Germany GE Healthcare UK Ltd Amersham Place Little Chalfont Buckinghamshire, HP7 9NA, UK GE Healthcare Bio-Sciences Corp 800 Centennial Avenue P.O. Box 1327 Piscataway, NJ , USA GE Healthcare Bio-Sciences KK Sanken Bldg , Hyakunincho Shinjuku-ku, Tokyo , Japan Asia Pacific Tel: Fax: Australasia Tel: Fax: Austria Tel: Fax: Belgium Tel: Fax: Canada Tel: Fax: Central & East Europe Tel: Fax: Denmark Tel: Fax: Eire Tel: Fax: Finland & Baltics Tel: Fax: France Tel: Fax: Germany Tel: Fax: Greater China Tel: Fax: Italy Tel: Fax: Japan Tel: Fax: Korea Tel: Fax: Latin America Tel: Fax: Middle East & Africa Tel: Fax: Netherlands Tel: Fax: Norway Tel: Fax: Portugal Tel: Fax: Russia, CIS & NIS Tel: Fax: Spain Tel: Fax: Sweden Tel: Fax: Switzerland Tel: Fax: UK Tel: Fax: USA Tel: Fax: AB 09/06 Printed in Sweden.