|
|
Bio Chromatography
FPLC- Fast Protein Liquid Chromatography System
GE Amersham Pharmacia ÄKTApilot
(AKTAP)
The ÄKTApilot is a dedicated automated liquid chromatography system designed for process development, process scale-up, and production. The system has the capacity to purify from milligrams to tens of grams of product and is bio-compatible, hygienic and sanitizable.
Q: Why is this unit the 'gold standard' ?
A: Because ÄKTApilot meets all good laboratory practice (GLP) and current good manufacturing practice (cGMP) demands for Phase I–III in drug development and final-scale production.
A new system retails for approximately $135,000 from GE Life Sciences, but from GMI you can buy a tested re-certified system for much less.
ÄKTApilot is a complete system and contains all the fluid handling components required to perform fluent delivery and mixing, sample application, and in-line monitoring. All the control and mechanical components are located inside an easily cleaned cabinet.
The flow path consists of two sets of four inlets on the main pump and four inlets on the sample pump. The latter has the same capacity as the system pump, 400 ml/min. The in-line mixer ensures high accuracy of the high-pressure gradient.
The air trap is connected in parallel for easy actuation. It is possible to use two columns in series or one column at a time. Pressure is monitored at four points and it is possible to pressure regulate the flow rate for convenient sample application and column packing. Monitoring of conductivity, UV and pH are all displayed in UNICORN and extra monitors can be connected to the system for special purposes.
Fractionation is performed either by the standard setup of eight outlet valves or by adding external valve blocks.
A fraction collector is typically not supplied as most chromatographers simply collect their fractions into sterile 'buckets' !
ÄKTApilot is equipped with sanitarily designed sensors for continuous in-line measurement of pH, conductivity and UV absorbance. The detectors provide accurate and reliable monitoring through self-test and self-calibration. The flow cells are placed close together, which minimizes band broadening and time delay between the detectors. The flow cells are easily accessible from the front panel to facilitate maintenance.
The UV cell can measure up to three wavelengths simultaneously, in the range 190–700 nm, via fiber optics. The pH electrode is mounted in the pH fl ow cell after pressure sensor 4 in the fl ow path. The pH monitor measures in the range 2–12. The conductivity cell is placed after the pH electrode in the fl ow path. It is used to verify gradients, follow peak positions relative to ionic strength and measure temperature via a built-in sensor.
| UV flow cell specifications | |
| Path length | 1 mm, 2 mm, 5 mm |
| Maximum pressure | 2 MPa (20 bar, 4.2 psi) |
| Maximum back pressure | 0.03 MPa (0.3 bar, 4.2 psi) |
| Wavelength accuracy | ± 2 nm |
| Wavelength accuracy | ± 2 nm |
| Wavelength precision | ±0.01 nm |
| Hold up volume | 300 µl |
| pH flow cell specifications | |
| pH range | 0-14 (valide between 2 & 12) |
| Response time | maximum 10 s (0-95% of step) |
| Maximum pressure | 0.5 MPa (5 bar, 72 psi) |
| Maximum back pressure | 0.01 MPa (0.1 bar, 1.4 psi) |
| 0.01 MPa (0.1 bar, 1.4 psi) | 240µl |
| Conductivity cell specifications | |
| Conductivity range | 1µS/cm to 999.9 mS/cm |
| Deviation from theoretical | Maximum ±2% of full scale range or conductivity ±10 µS/cm whichever is greater in the range 1µS/cm to 300mS/cm |
| Response time | maximum 3s (0-95% o f step) |
| Temperature sensor accuracy Drift | ±2.0°C ±0.5°C per 10 h |
| Maximum back pressure | 0.02 MPa (0.2 bar, 2.9 psi) |
| hold up volume | 180 µl |
| Mixer specifications | |
| maximum back pressure | 0.05 MPa (0.5 bar, 7 psi) |
| mixing principle | chamber with a stirrer |
| hold up volume | 4.2 ml |
| Pressure sensor specifications | |
| maximum pressure | 2.5 MPa (25 bar, 363 psi) |
| maximum back pressure | 0.06 MPa (0.6 bar, 9 psi) |
| hold up volume | 331µl 2-port 346µl 3-port |
| Pump & sample pump specifications | |
| flow rate isocratic and gradient mode isocratic and limited gradient | 4-400 ml/min up to 800 ml/min |
| pressure | 0-2 MPa (20 bar, 290 psi) |
| flow rate accuracy | < ±2% (4-400 ml/min, 0.1-2 MPa) |
| flow rate reproducibility | rsd < 0.5% (4-400 ml/min, 0.1-2MPa) |
| viscosity | maximum 5 cP |
| Gradient composition accuracy | ±3% at 8-400 ml/min |
| Hold up volume | Â |