The AKTAPurifier 10 : Ideal for Smaller Purification Quantities

The AKTA Purifer 10 If your applications demand a wide variety of wavelength, then consider the AKTA Purifier 10 system. The tunable detector featured in this model allows you to select from a wide spectral range. The AKTA Purifier 10 allows of flow rates of up to 10 ml per minute.

AKTApurifier 10 UPC: For Filter Based Wavelength Detection

The AKTA Purifier 10 UPC The AKTA Purifier 10 UPC is the most economical choice of the 4 units. It is ideally suited for applications which require up to 10 ml of flow rate per minute and where a filter based wavelength detector is adequate.

The AKTAPurifier 100 : Ideal for Larger Purification Quantities

The AKTA Purifier 100 If your applications demand a wide variety of wavelength, then consider the AKTA Purifier 100 system. The tunable detector featured in this model allows you to select from a wide spectral range. The AKTA Purifier 100 allows of flow rates of up to 100 ml per minute.

AKTApurifier UPC 100: For Filter Based Wavelength Detection

The AKTA Purifier 100 UPC This is the most economical choice of the 4 units. It is ideally suited for applications which require up to 100 ml of flow rate per minute and where a filter based wavelength detector is adequate.

Introduction to the GE AKTA Purification System

The GE AKTA Purifier (ÄKTApurifier™) is a liquid chromatography system designed for fast and reliable separations of peptides, nucleic acids, oligonucleotides and proteins. ÄKTApurifier™ runs all techniques, from μg to mg scale at flow rates 0.001 m/min - 10 ml/ min and pressures 0-25 MPa. With operational simplicity and a wide range of pre-programmed application protocols and method templates, ÄKTApurifier™ incorporates many features to minimise preparation and run times and maximise reliability and reproducibility of results. ÄKTApurifier™ is essential to guarantee fast, reproducible runs everyday.

 

The GE AKTA Purifier, Fast and easy start to purification work

With the AKTA Purifer, users are now able to create complete purification schemes for a range of biomolecules. Application protocols and method templates combine to create and run chromatographic methods. This makes ÄKTApurifier™ extremely easy to use and speeds up routine purifications. It reduces the need for expertise in programming, in chromatography and in time consuming literature searches, as all this knowledge is built into ÄKTApurifier™. Application protocols and Method templates All protocols and templates are based on extensive, well documented work in our laboratories, and collaboration with experts.

Protocols in ÄKTApurifier™ include:

  Purification of synthetic peptides Purification of labelled synthetic oligonucleotides Purification of native peptides Purification of protein fragments Methods can be created easily in less than 3 minutes.   Â

Each Method Template contains certain parameters as variables, e.g. flow rate, sample volume, column equilibration, for simple changes before each run if required. Pre-programmed Method Templates eliminate time consuming and repetitive method programming. The templates use column volume as a programming base for easy use with any size of column. Time base programming is also available. User specific templates can be created easily

Standard Method Templates include:

1. Basic templates: linear gradients as appropriate

• Size exclusion chromatography
• Anion exchange chromatography
• Cation exchange chromatography
• Hydrophobic interaction chromatography
• Reversed phase chromatography
• Afinity chromatography

2. Optional templates

• Manual sample application via sample loops or SuperJoops (includes repeat injection option)
• Automatic sample application via system pump
• Segmented gradient elution
• Fraction collection by fixed volume or automatic peak fractionation
• Speed and reliability for every run
• Fast routine runs
• To quickly establish fast routine runs, easy to use templates for method scouting are included
• These templates make it easy to systematically vary run parameters in repeated, automated procedures, reducing optimization times.

Fast peak identification with the ÄKTApurifier™

User selected multi-wavelength monitoring enables on-line detection of specific labelled products and can be used to detect contaminants at higher sensitivity wavelengths (e.g. 280/2141254 nm).

Accurate result comparisons

All data, including running conditions, are stored permanently for every run within the run log e.g. flow, pressure, UV, conductivity, temperature. This makes it extremely easy to compare runs and determine a cause for any deviations which may be seen.

Accurate peak fractionation - improve peak purity

Accurate peak fractionation can help to improve the purity of a selected peak. Peak fractionation can be based on peak detection using slope or level sensing. Fraction marks and fraction numbers make it easy to identify fractions and peaks.

Minimal procedure times User selected and defined WATCH functions (see Figure 9) can be entered into a method template to instruct specific events to occur when preset conditions are reached. For example, the WATCH function will wait for a return to normal baseline level before instructing the next run to begin, minimising the time spent on column re-equilibration between runs.

Column knowledge - built-in

ÄKTApurifier™ ensures every column is run optimally and is protected by ÄKTApurifier™ from damaging events such as over-pressure. The user selects a pre-packed column from the extensive column library and ÄKTApurifier™ automatically sets the optimal running parameters for that column (e.g. column volume, flow rate and pressure limit) into the method. The user can adjust running conditions (if required) by simple method editing. New columns can be added to the library and will be run in the same secure way.

The selection of the correct column for each purification step is essential for optimum results. The Mini Q can be used for improved resolution. The Mono Q yields a higher purity product after only a single step. Optimum elution and accurate peak fractionation in ÄKTApurifier™ ensure a minimum number of purification steps.

Increased reliability and reproducibility from day to day
Accurate, automatic buffer preparation

The unique function, BufferPrep, eliminates the time-consuming manual buffer preparation and titration needed for every pH change in chromatography.

The user selects the buffers to be used from the buffer library and defines the pH required. BufferPrep prepares correct buffer pH on-line from four stock solutions (e.g. water, salt, buffer, acid/base) and controls the pH throughout the run, automatically compensating for changes in temperature and salt concentration. New buffer substances can be added to the buffer library and used by BufferPrep to create the required elution pH conditions.

The accuracy of pH is crucial in many separations. The accuracy of BufferPrep greatly increases the reproducibility and reliability of results from day to day by eliminating and correcting errors which occur when producing buffers manually. BufferPrep can be used for pH scouting in rapid method optimisation. Pre-programmed method templates are available for automatic eluent pH scouting for ion exchange chromatography. Figure 15 shows BufferPrep used on AKTAexplorer at 60 mis/min to test seven pH conditions to find the optimal pH for elution of a pure Fab fragment.

Automatic evaluation and reporting

For convenience ÄKTApurifier™ includes pre-programmed evaluation and report procedures which can be included within run methods. User- defined evaluation procedures and customised report layouts can also be quickly created.

System description General description of the ÄKTA purifier™.

ÄKTApurifier™ has three main instrument components, a number of ancillary items, and the option of a fraction collector and autosampler. The positions of the main components are indicated in the diagram. All instrument settings and functions are under direct control of UNICORN.

At every start-up, each instrument automatically performs a self-diagnostic test and calibrates appropriate settings. This ensures top performance throughout day-to-day operation.

All wetted materials are inert, fully biocompatible and resistant to all eluents commonly used in liquid chromatography, ensuring high recovery and no loss of biological activity.

The system is supplied assembled and tested. The system is mounted on a swivel platform and uses minimal bench space. Any side can be turned to face the operator for easy accessibility. Columns are easily snapped in place on the outside of the system.

A simple and logical flow configuration of ÄKTApurifier™ system includes:

• easy pump wash for quick solvent exchange without disturbing the column
• automatic on-line buffer preparation from stock solutions
• automatic sample application - fixed volume loops from 100ml to 1 ml, volumes smaller than 100 ml are applied by partial loop filling.
  - SuperJoop'M 10 ml, Superloop 50 ml and SuperJoop 150ml for the range 1-10 ml, 1-50 ml and 1-150 ml respectively.
  - larger volumes via the system pump, Pump P-903, for the application of several litres of sample.
• on-line UV-detection of up to three wavelengths simultaneously
• on-line monitoring of conductivity
• optional on-line monitoring of pH
• optional collection of up to 175 fractions
• routine, automated system wash.

System control - UNICORN

UNICORN is a control system for real-time control of AKTAdesign systems. UNICORN as a control platform ensures quick and simple communication between all AKTA systems and AKTA users. UNICORN control system can be used from laboratory to full scale production. The same interface and methods used to run routine applications can be used to optimize a purification in research and development and applied at production scale. UNICORN software is installed on the PC prior to delivery and controls all AKTAdesign instrument settings and functions. For further information on the latest version of UNICORN system control, please contact your sales representative.

System components

Pump P-901 and P-903 (P-900 series pumps deliver accurate, reproducible flow rates and gradients for all biomolecular chromatographic separations).

Pumps P-901 (up to 100 ml/min) and P-903 (up to 10 ml/min) employ features that allow all modern high-performance media to be run under optimal conditions and maximize the results of every chromatographic separation. The key features include the use of four pump heads instead of two. This permits accurate flow and gradients over a wide range of flow rates and the incorporation of three microprocessors to monitor and control performance.

The result is high accuracy and reproducibility, automated monitoring of performance and safety, column protection through the use of gradual starts and pressure limits, and an internal log for easy user monitoring.

Monitor pH/Cond-900 (Monitor pH/C-900 is a combined monitor for accurate, precise online monitoring of pH and conductivity conditions over a wide dynamic range).

Monitor pH/C-900 is a combined monitor for accurate, on-line monitoring of pH, conductivity and temperature recording real time information about chromatographic conditions.

Its accurate response, coupled with high precision over a wide measuring range, makes it ideal for use in all chromatography techniques, from reversed phase with very low conductivity eluents to hydrophobic interaction chromatography in high salt solutions.

The pH/C-900 consists of a control unit, a flow cell for conductivity and temperature, a flow cell with a holder for the pH electrode and the pH electrode.

Monitor UV-900 (Monitor UV-900 combines high sensitivity UV monitoring at three wavelengths simultaneously).

Monitor UV-900 is not only an integral component of ÄKTA™design chromatography systems, but is also an excellent standalone multi-wavelength detector. The application of fiber technology to the design of the flow cell means that focused light is continually reflected back into the flow path. The cell features a long path length while retaining a small volume and a design that prevents formation of lens-shaped interfaces between eluant components of different refractive indexes. Fiber optics allow the flow cell to be placed close to the column outlet. These innovations increase signal-to-noise ratio, while decreasing band broadening, noise, and thermal drift. The result is a very sensitive, stable, and reliable UV/visible detector.

Monitor UV-900 is a high sensitivity multi-wavelength UV-Vis monitor, recording up to three wavelengths simultaneously for optimal detection in the range 190-700 nm.

The use of advanced fiber optics together with the unique flow cell design ensures a high signal-to noise ratio for maximum performance. Maintenance costs are minimized by use of a unique Xenon flash lamp, giving a significant increase in lamp lifetime.

Monitor UV-900 includes a choice of two flow cells (optical path-length 2 mm, internal volume 2μl, or path-length 10 mm, internal volume 9 μl). The 10 mm flow cell is delivered with ÄKTApurifier™

Monitor UPC 900 (Monitor UPC-900 - A single unit measures UV, pH, and conductivity)

Monitor UPC-900 provides UV and temperature-compensated pH and conductivity parameters in a single display. Temperature is monitored and displayed and can be recorded if the monitor is used in ÄKTA design systems.

The mercury lamp is available with 254 or 280 nm filters for detecting proteins; the zinc lamp is used with the 214 nm filter for detecting peptides or for increased protein detection sensitivity. Additional filters are available for special applications within the range of 313 to 546 nm. The 2 mm UV flow cell and the pH and conductivity flow cells are designed to accommodate flow rates up to 100 ml/min; the 5 mm UV flow cell is optimized for flow rates to 20 ml/min.

The conductivity monitor features a broad dynamic range which accommodates the high salt concentrations used in hydrophobic interaction chromatography and the low conductivity of reversed phase or desalting separations. The pH monitor is essential for optimal results in affinity, ion exchange, and chromatofocusing separations.

Ancillary equipment: standard

ÄKTApurifier™ is supported by a full range of dedicated ancillary equipment. All ancillary equipment is controlled via UNICORN control system.

Valves ÄKTApurifier™ has one seven port injection valve, INV-907, for sample application and one eight port fractionation valve, INV-908, for diversion to fraction collector or waste. Two switch valves work together with the P-903 pump in BufferPrep mode.

Mixer M-925

P-900 series pumps deliver accurate, reproducible flow rates and gradients for all biomolecular chromatographic separations.

Mixer M-925 is a dynamic, single chamber mixer powered and controlled from Pump P-903.

For optimal mixing in the entire flow rate range of ÄKTApurifier™, it is delivered with interchangeable mixing chambers (0.6 and 2 ml).

 

Ancillary equipment: optional

Fraction Collector FRAC-901 FRAC-901 can be used for both small and preparative scale purifications with ÄKTApurifier™. It collects up to 175 fractions in 12 mm diameter tubes or up to 95 fractions in 10-18 mm diameter tubes.

In ÄKTApurifier™, the FRAC-901 allows fixed volume fractionation or automatic peak fractionation. Peak fractionation can be based on peak detection using slope or level sensing. Fraction marks and fraction numbers make it easy to identify fractions and peaks.