Äkta Design

ÄKTAdesign FOR more information ABOUT One platform
THE COMPLETE ÄKTAdesign RANGE,

WHAT TYPE OF WORK GO TO THE ÄKTA HOMEPAGE AT: simplifies ALL YOUR
ARE YOU DOING? www.chromatography.amershambiosciences.com PURIFIC ATION TASKS
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What type of purification is And you’ll save time, because ÄKTAdesign allows everyone
Tel: +852 2811 8693 Tel: 0169 35 67 00 Tel: 21 417 7035
going on in your lab? to work in the same, logical fashion. ÄKTAdesign comes Fax: +852 2811 5251 Fax: 0169 41 9677 Fax: 21 417 3184
Are you purifying proteins for structural or functional
studies? Are you developing protocols and optimizing
with a method wizard to facilitate programming, automatic
buffer preparation, and a complete range of prepacked
Australasia
Tel: +61 2 9899 0999
Fax: +61 2 9899 7511
Germany
Tel: 0761 4903 401
Fax: 0761 4903 405
Russian & other C.I.S. & N.I.S.
Tel: +7 (095) 232 0250, 956 1137
Fax: +7 (095) 230 6377
ÄKTAdesign
methods for biomolecule purification? Are you purifying columns. Whatever your application or scale of purification,
Austria Italy South East Asia
synthetic peptides or nucleic acids? you will have the fastest, simplest approach to purification. Tel: 01 576 0616 20 Tel: 02 27322 1 Tel: 60 3 8024 2080
With ÄKTAdesign, you can perform any type of purification Fax: 01 576 0616 27 Fax: 02 27302 212 Fax: 60 3 8024 2090
Regardless of whether you work in the field of.... with speed, ease, and flexibility. Belgium Japan Spain
• Structural studies Tel: 0800 73 888 Tel: 81 3 5331 9336 Tel: 93 594 49 50
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• Functional studies Latin America
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• Process development for Windows 2000/NT: no other control Tel: 1 800 463 5800 Tel: +55 11 3667 5700 Tel: 018 612 19 00
....you need to purify proteins and there is an system is this user friendly. Fax: 1 800 567 1008 Fax: +55 11 3667 87 99 Fax: 018 612 19 10
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ÄKTA™design system for your needs. Conducting and controlling purification tasks couldn’t be Tel: +30 (1) 96 00 687
South East Europe Tel: 01 802 81 50
simpler. From planning to final report, from lab to production Tel: +43 1 982 3826 Fax: +30 (1) 96 00 693 Fax: 01 802 81 51
If your focus is fast method and process development
scale, UNICORN™ for Windows™ 2000/NT™ gives you Fax: +43 1 985 8327 Netherlands UK
ÄKTAexplorer is ideal, while ÄKTApurifier is designed Tel: 0165 580 410
choices all the way. Denmark Tel: 0800 616 928
for those who routinely purify and characterize proteins, Tel: 45 16 2400 Fax: 0165 580 401 Fax: 0800 616 927
peptides, and nucleic acids. ÄKTAFPLC™ is optimized for Need to modify your system configuration? No problem. Fax: 45 16 2424 Norway USA
Finland & Baltics Tel: 2318 5800 Tel: +1 800 526 3593
research and development with proteins. And if you New flexible system software lets you add new functions Fax: 2318 6800
Tel: +358 (0)9 512 3940 Fax: +1 877 295 8102
need a compact, ready to use, push-button solution for and UNICORN will do the rest. Fax: +358 (0)9 512 1710
simple protein purification, ÄKTAprime is the answer.
As you scale-up your work, UNICORN ensures that your
One platform from basic research pilot- and production-scale protocols will operate as
to process development. smoothly as your initial purification strategies.
Äkta is the Swedish word meaning true; genuine; real. With unique networking capabilities, UNICORN gives you ÄKTA, UNICORN, FPLC, SOURCE, RESOURCE, Sephasil, Superdex, HiPrep, GSTrap, HiLoad,
MiniBeads, BufferPrep, Sepharose, HyperD, Superloop, HiTrap and Drop Design are trademarks
ÄKTAdesign is the name for a platform of systems, total control—from your desk. of Amersham Biosciences Limited. Amersham and Amersham Biosciences are trademarks of
control software, and columns that help you make real Amersham plc.
Amersham Biosciences AB Björkgatan 30, SE- 751 84 Uppsala, Sweden.
progress in biomolecule purification. Amersham Biosciences Amersham Place, Little Chalfont,
Buckinghamshire HP7 9NA, England.
Amersham Biosciences Corp 800 Centennial Avenue, PO Box 1327,
Piscataway, NJ 08855 USA.
Amersham Biosciences Europe GmbH Munzinger Strasse 9,
D- 79111 Freiburg, Germany.
Amersham Biosciences Sanken Building, 3-25-1, Shinjuku-ku,
Tokyo 169-0073, Japan.
All goods and services are sold subject to the terms and conditions of sale of the company 18-1158-77
within the Amersham group that supplies them. A copy of these terms and conditions is
available on request. © Amersham Biosciences AB 2001 - All rights reserved. Edition AA
Tel: +852 2811 8693 Tel: 0169 35 67 00 Tel: 21 417 7035
Australasia
Tel: +61 2 9899 0999
Fax: +61 2 9899 7511
Germany
Tel: 0761 4903 401
Fax: 0761 4903 405
Tel: +7 (095) 232 0250, 956 1137
Fax: +7 (095) 230 6377
ÄKTAdesign
Canada Sweden
Tel: +358 (0)9 512 3940 Fax: +1 877 295 8102
ÄKTAprime UNICORN FOR WINDOWS 2000/NT. HOW TO ADD EVEN
simple purification of proteins.
ANY PURIFIC ATION TASK. more flexibility
ÄKTAprime is ideal whenever, and wherever,
a simple, rapid, protein purification is needed in
EVERY PURIFIC ATION SC ALE. TO YOUR SYSTEM
the laboratory. Compared to traditional manual
operations, ÄKTAprime offers significant
advantages in speed and capacity.
ONE FASTER, more intelligent Frac-901
No matter whether the task is a quick sample

clean-up or a purification from a large volume WAY OF WORKING.
of cell culture supernatant, this compact system
will be ready to use. Supplied as standard with all computer-controlled systems offers functions that are vital in today’s R&D laboratories and
in the ÄKTAdesign family, UNICORN gives you a common production facilities. Method wizard and scouting functions
With pre-installed application templates and
control platform and one common user interface for all simplify purification tasks, while the ability to generate
prepacked columns, ÄKTAprime performs the
scales of operation. This unique control system software customized repor ts and documentation saves time. UNICORN
most common purification steps at the touch Frac-950
with its networking capabilities puts you in total control is also supplied with comprehensive documentation that helps
of a button.
throughout your facility—from lab-scale research to full- to fulfil regulatory requirements. UNICORN complies with
scale production. From planning to final report, UNICORN FDA 21 CFR par t 11 which also includes electronic signature. Fraction
collector Autosampler
Simple Flexibility Sample

pump
A/D
PrimeView
Automated Wizard multiple
converter
Air
sensors
peak compare. On line
pH
Flexibility
flowpath
Column scouting
Method wizard.
Reversed flow
Buffer selection
PrimeView software allows for continuous real-time monitoring of the separation process and provides facilities
for presentation and evaluation of separation results. Data output.
Standard configuration ÄKTAFPLC ÄKTApurifier ÄKTAexplorer
high performance purification high performance purification fast method and process development
ÄKTAexplorer 100 Air ÄKTApurifier XT of proteins. and characterization of proteins, and scale-up for proteins, peptides,
peptides, and nucleic acids. and nucleic acids.
Buffer Selection Air Sensor Flow Direction
BufferPrep A B Autosampler A-900
BufferPrep A2 A1 C D ÄKTAFPLC builds on the world-wide success of

B1 B2 Injection Mixer FPLC™ System, but offers major advantages in For fast, reliable separations of peptides ÄKTAexplorer is designed for all
valve Pump
Mixer Injection valve terms of design and versatility. and nucleic acids at laboratory scale, chromatographic techniques employed
Pump
ÄKTAFPLC offers the choice between one-step ÄKTApurifier is unbeatable. It is equally at in method and process development,
Multiple Sample pH, Cond UV-Vis
Injection Column Scouting procedures or automated multi-column home with routine procedures of any offering microgram to gram scale
Sample Pump
purification schemes to purify proteins in complexity, with scouting and method purification. Developing optimized
Air Sensor
pH, Cond UV-Vis Fractionation microgram to milligram scale at flow rates optimization, and with biomolecule methods is fast and easy. The scouting
up to 20 ml/min. characterization, such as peptide mapping. function of UNICORN automatically
varies parameters over repeated runs.
The innovative, compact design makes the The system performs all chromatographic
Fractionation BufferPrep calculates and prepares the
system extremely easy to work with, without techniques, helping you to scout for optimal
correct pH buffers from stock solutions,
losing flexibility. The integral rack opens to conditions for binding and elution, pH,
while automatically compensating for
accommodate any column. gradient shapes, and flow rates. The method
ÄKTA FPLC ÄKTAprime changes in temperature and salt
wizard provides a unique method and
concentration.
Buffer and sample selection suppor t for all chromatographic techniques.
Buffer A
Mixer
Buffer A
Buffer B
Injection Valve
Pump NO
Buffer B NC
Gradient Mixer
Manual injection or via Superloop Pump Injection valve
formation
valve
UV and Cond, Manual injection or via Superloop
(pH optional)
UV and Cond,
(pH optional)
Diversion to waste
Fractionation
Diversion to waste
Fractionation multistep
Way of working
Simple, one-step purification
Standard ÄKTAdesign configurations
Explorer
!
Purifier
!
FPLC
!
Prime
!
Buffer Prep
Reproducible performance for
! ! ! !
routine purification
Optimization of one-step purification
to increase purity
System control and data handling for
regulator y requirements, e.g. GLP
!
!
!
!
!
!
! Dedicated
configurations for
Characterization and analysis Scouting
Automatic method development
and optimization ! ! !
Automatic buffer preparation ! ! structural genomics The same, tried-and-tested pump technology
Automatic pH scouting
Automatic media or column scouting
Automatic multi-step purification
Scale-up, process development, and
transfer to production
!
!
!
!
!
soon to come.....
The integral rack of ÄKTAFPLC opens to
that you’ll find in FPLC—but now with twice
the throughput.
- Non air sensitive.
- High salt resistant.
Multidimensional purification
allow a wide variety of configurations. - Transparent.

valve Pump
Pump
Sample Pump
Air Sensor
concentration.
Buffer A
Mixer
Buffer A
Buffer B
Injection Valve
Pump NO
Buffer B NC
Gradient Mixer
formation
valve
(pH optional)
UV and Cond,
(pH optional)
Diversion to waste
Fractionation
Diversion to waste
Way of working
Explorer
!
Purifier
!
FPLC
!
Prime
!
Buffer Prep
! ! ! !
to increase purity
!
!
!
!
!
!
! Dedicated
configurations for
!
!
!
!
!
soon to come.....
the throughput.

valve Pump
Pump
Sample Pump
Air Sensor
concentration.
Buffer A
Mixer
Buffer A
Buffer B
Injection Valve
Pump NO
Buffer B NC
Gradient Mixer
formation
valve
(pH optional)
UV and Cond,
(pH optional)
Diversion to waste
Fractionation
Diversion to waste
Way of working
Explorer
!
Purifier
!
FPLC
!
Prime
!
Buffer Prep
! ! ! !
to increase purity
!
!
!
!
!
!
! Dedicated
configurations for
!
!
!
!
!
soon to come.....
the throughput.
Capture and isolation of fusion proteins and antibodies Purification of proteins, antibodies, and enzymes Peptide mapping, analysis, and characterization Scouting, method development, and scale-up of
Capture and isolation of fusion proteins and antibodies is a task best Working with routine purification of molecules like proteins, antibodies, Mapping or characterizations of proteins, peptides, or nucleic acids biomolecule purification
w
Flo
Sample: Cell culture supernatant containing mouse IgG1 Capture Column size: 1 ml, 5 mm x 50 mm
F
Column: SOURCE 5RPC ST 4.6/150
st
rD
and enzymes is slightly different to other purification tasks.
Fa
performed using affinity chromatography. Column: HiTrap Protein G HP 1 ml often require high-selectivity analytical methods. Reversed phase Start
pe
XL
Column: HiPrep 16/10 Q XL Sample: Synthetic Amyloid-b 1–42
e™
Hy
Q
Q
Binding buffer : 20 mM sodium phosphate, pH 7.0 buffer (A): 25 mM Tris-HCl,
e
15
30
os
os
Sample
Scaling-up methods for process scale, as well as defining
ic
Sample: Clarified E. coli extract 0.5 M NaCl,
chromatography (RPC) is often used in applications such as these.
ar
ar
CE
CE
Elution buffer: 0.1 M glycine-HCl, pH 2.7 amount: 622 µg
ra
ph
ph
1 mM EDTA,
Affinity chromatography methods typically use step gradients, which For purification of proteins, antibodies, and enzymes robustness and
UR
UR
Sample volume: 40 ml
Ce
250
Se
Se
ity
Eluent A: 10 mmol/l NH4OH-HCOOH, pH 6.4, 8.2 and 9.0 respectively pH 8.0
parameters for a validated purification scheme or analysis
SO
SO
tiv
The requirements of low system dead volume and precise gradient
Q
Start buffer (A): 50 mM Tris-HCl, 1 mM EDTA, pH 7.5; 2 mM DTT,
Absorbance 260 nm
uc
Eluent B: 60% acetonitrile Elution
are fairly simple to perform and do not require the use of complex reproducibility are most impor tant provided that the selectivity is good 0.2 M benzamidine-HCl, 0.2 mM PMSF 200
nd
buffer (B): 25 mM Tris-HCl,
method, all require optimization.
Co
Flow rate: 0.5 ml/min
AU 280 nm pH Elution buffer (B): A + 1.0 M NaCl accuracy place high demands on the purification system, par ticularly 1.2 M NaCl, 150
instrumentation. enough. Being sure that the system will be able to work around the clock Gradient: 0% B in 5 CV, 30% B in 5 CV, 100% B in 5 CV (step gradient) Gradient: 20–80% B over 87.5 ml (35 CV) 1 mM EDTA,
UV 280 nm Flow rate: 10 ml/min (300 cm/h) with organic solvents as used in RPC. Detection: 230 nm
Regardless of target molecule, scouting of separation
pH 8.0 100
pH
with a minimum of maintenance is crucial. Sample: 76 µg of
Instead of tying up a high-end chromatography system, a simple, mAU mS/cm
NH3 -HCOOH/CH3CN, 60% 4.5 kb plasmid
50
1.0
When working with these kinds of applications, it is an advantage if conditions and scouting for the right column is essential to Gradient: 0–60% B, 30 CV 0
dedicated system can be used for the task. A purification instrument with a pump system that is not sensitive to air 3000 mAU
7 the purification system used is targeted for the specific needs. A 230nm
230 nm
%B finding the best possible method parameters. 10.0 15.0 20.0 25.0 30.0 35.0 ml
and salt is highly valued for routine purification. DAOCS (mAU) Elution of plasmid on different anion exchange media.
2000 254 nm
30.0
Also, high flexibility in combining and running several methods
280 nm 80
6
Sample: Detergent extracts of E. coli membranes Column: Superdex™ 200 HR 10/30
Column: Sephasil™ Peptide mAU
in series is often needed if high purity is a requirement. Sample: 10 ml clarified cell homogenate Intermediate step: Desalting
Binding buffer: 20 mM Tris-HCl, pH 7.5, with 5 mM imidazole, Sample 1000
C18, 5 µm ST 4.6/250 pH 6.4 containing His-GFP
0.8
0.03% dodecyl-b-D-maltoside and application: 300 µl applied via 0.1 ml Column: HiPrep 26/10 Desalting
300 mM NaCl sample loop Sample: Tr yptic digest RVP ovalbumin 20.0 60 A chromatography purification system that meets these Step 1: Affinity chromatography
Buffer : 20 mM Tris-HCl, pH 8.0
Elution buffer (B): 20 mM Tris-HCl, pH 7.5, with 500 mM imidazole, Buffer: 20 mM Tris-HCl, pH 7.5, with 0
Flow rate: 1 ml/min Column: HiTrap Chelating HP 5 ml,
Sample: Clarified homogenate of E. coli expressing GST fusion protein Flow rate: 10 ml/min
Column: GSTrap™ FF 1 ml 5
0.03% dodecyl-b-D-maltoside and 1.5 % octyl-b-D-glucoside,
0 100 200 ml
Eluent: CH3CN/TFA 100
needs will shor ten the time needed to complete the charged with Ni2+
300 mM NaCl 150 mM NaCl Step 2: Ion exchange
Binding buffer : 20 mM Tris-HCl, 0.5 M NaCl,
Binding buffer : 20 mM sodium phosphate, 0.15 M NaCl, pH 7.3 Gradient: 0–60% B, 20 column volumes Flow: 0.25 ml/min 10.0 40 method optimization. 20 mM imidazole, pH 7.4 Column: RESOURCE Q 6 ml
Elution buffer (B): 50 mM Tris-HCl, 10 mM glutathione, pH 8.0 Temperature: 5 °C Temperature: 5 °C
Mouse Intermediate purification 50 Elution buffer : 20 mM Tris-HCl, 0.5 M NaCl, Binding buffer : 20 mM Tris-HCl, pH 8.0
System: ÄKTAFPLC System: ÄKTAFPLC 0.5 M imidazole, pH 7.4
IgG 1 4 Column: SOURCE™ 15ISO, packed in Elution buffer : 20 mM Tris-HCl, 1.0 M NaCl,
Sample loop: Superloop™ 10 ml or 50 ml Flow rates: Binding, wash and equilibration pH 8.0
0.4 HR 16/10 column (16 mm x 50 mm bed) 0.0 20
Sample: Fab fraction from Gradient: 20 column volumes, to 1 M NaCl
UV 280 nm %B Sample volume: 8.5 ml 0 10 ml/min, elution 5 ml/min
AU 280 nm HIC separation, 20 ml Flow rate: 60 ml/min Flow rate: 10 ml/min
Programmed %B Sample: DAOCS pool from HiPrep 16/10 Q XL
Column: HiTrap Chelating HP 1 ml, Column: RESOURCE S 6 ml
mAU mS/cm Sample volume: 40 ml Curves: A280 nm, from top:
charged with Ni2+ Eluate from Eluate from IEX column
1.6 Inject Start buffer (A): 1.6 M ammonium sulfate, 10% glycerol, 50 mM Tris-HCl, Eluents: Automatic BufferPrep™ with pH 4.0; 4.5; 5.0; 5.5; 6.0; 6.5; 7.0.
3 Flow rate: 1 ml/min 60 0.06 M sodium acetate, A280 Eluate from affinity desalting column collected in fraction collector
10 1 mM EDTA, 2 mM DTT, 0.2 mM benzamidine-HCl, 0 20 40 60 80 100 120 ml 0 System: ÄKTAexplorer
80 0.03 M sodium phosphate, mAU column collected collected in Loop 2
0.2 mM PMSF, pH 7.5 0.0 20.0 40.0 60.0 ml
mA U %B 0.03 M sodium formiate, in Loop 1
Elution buffer (B): 50 mM Tris-HCl, 10% glycerol, 1 mM EDTA, 2 mM DTT, High reproducibility—essential for peptide mapping. 0.1 M HCl and 2 M NaCl
8 0.2 mM benzamidine-HCl, 0.2 mM PMSF, pH 7.5 2000
2 100 Gradient: 0–16% B in 4 CV, 16–24% B in 8 CV, 24–35% B in 4 CV,
0 START CONDITIONS Run 1 Run 2 Run 3 Run 4 Run 5 Run 6 Run 7
60 100% B in 4 CV A 230nm Elution
1500 6 %B pH 7 6.5 6 5.5 5 4.5 4 Salt peak Re-equilibration
80 40 Flow rate: 5 ml/min (150 cm/h) 230 nm of IEX column
gradient
7.5 15.0 min (mAU) mAU 280 nm 1500
0.8 Column: Sephasil Peptide mAU pH 8.2 Conductivity (mS/cm)
Fraction 2 60 mAU mS/cm C18, 5 µm ST 4.6/250 254 nm
1000 4
GST 40 Fraction 1 DAOCS Sample: Tr yptic digest RVP ovalbumin 30.0 280 nm 80 80.0 A 280
fusion 40 1000
Flow rate: 1 ml/min 250
Monoclonal antibody purification. 2 400
protein 500 Eluent: CH3CN/TFA
20
20 20.0 500
20 300 60 200
0 0 60.0
Inject 0
Sample: Clarified homogenate of E. coli expressing His fusion protein 0 10 20 30 ml 0 5 10 15 20 ml
200
Column: HiTrap Chelating HP 1 ml charged with Ni2+
Purification on HiTrap Chelating HP. Sample: Fraction 1 from HiTrap
150 0 F8Waste F7 Waste F7 Waste F3 Waste 45 7 9 11 14 1719 2224 27 3032 3537 4042 45 48
Binding buffer : 20 mM sodium phosphate, 0.5 M sodium chloride, 10.0 40 pH 4.5 50 100 150 200 250 300 350 400 450ml
0 0 10 mM imidazole, pH 7.4 Chelating HP 1 ml. 100 40.0
Elution buffer (B): 20 mM sodium phosphate, 0.5 M sodium chloride, Chromatographic characterisation on 100
0 12.5 25 min 0.5 M imidazole, pH 7.4 Sample: 12 ml clarified cell homogenate Step 2: Gel filtration
Superdex 200 HR 10/30. 0
0 100 200 ml containing His-GST
60 ml 0.0 20
Column: HiLoad 16/60 Superdex 75
50 20.0
Step 1: Affinity chromatography prep grade
GST fusion protein purification. Metal affinity purification of membrane protein. Accuracy of automatic peak fractionation in ÄKTApurifier. Column: GSTrap FF 5 ml Buffer : 20 mM HEPES,
Polishing
Binding buffer : 20 mM Tris-HCl, 0.15 M NaCl, pH 7.4 0.15 M NaCl, pH 7.4
%B Column: HiLoad 16/60 Superdex 75 prep grade 0 0
AU 280 nm 0.0 20.0 40.0 60.0 ml Elution buffer : 20 mM Tris-HCl, 0.15 M NaCl, Flow rate: 1.5 ml/min
0.3 100 Sample: Concentrated DAOCS pool from SOURCE 15ISO 10 mM reduced glutathione, pH 8.0
UV 280 nm 2.0 4.0 6.0 8.0 Time (min)
Recommended columns Programmed %B Recommended columns Sample volume: 3 ml
Recommended columns Flow rates: Binding and elution 1 ml/min,
Buffer: 100 mM Tris-HCl, 1 mM EDTA, 2 mM DTT, wash and equilibration 10 ml/min
0.2 mM benzamidine-HCl, 0.2 mM PMSF, pH 7.5 Automatic pH scouting on RESOURCE S 6 ml using A 280
A 230nm mAU
• HiTrap™ 80 Flow rate: 1 ml/min (30 cm/h)
230 nm
%B BufferPrep.
• High Resolution (HR) • SOURCE ST/PE (mAU)
mAU 254 nm 2000
pH 9.0
• HiPrep™ 0.2 • HiLoad™ 1000 • MiniBeads™ PE
30.0 280 nm 80
His fusion 60 Recommended columns 1500 Eluate from affinity column Eluate from gel filtration
protein collected in Loop 1 collected in fraction collector
• RESOURCE™ IEX • RESOURCE 800
DAOCS • Superdex Peptide 20.0 60
600
• HiLoad 1000
40 400
0.1
200 10.0 40 • HiPrep 500
20 0
0 20 40 60 80 100 ml
• RESOURCE
0.0 20 0 Waste F8 Waste 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
Inject 20 40 60 80 100 120 140 160 ml
The complete, optimized purification scheme Typical results from two different automated two-step
0 0
for DAOCS using ÄKTAFPLC. 0 purification schemes.
0 45 65 min 0.0 20.0 40.0 60.0 ml
a) Capture using AIEX. a) Purification of His-GFP using affinity chromatography followed by
b) Intermediate purification using HIC. desalting and ion exchange chromatography.
c) Polishing using gel filtration. The true gradient b (arrow) 1–42 on
Method optimization of Amyloid-b b) Purification of His-GST using affinity chromatography followed
His fusion protein purification. (conductivity trace; red) is shown in a) and b). SOURCE 5RPC ST 4.6/150 by gel filtration.
Recommended system: ÄKTAprime Recommended system: ÄKTAFPLC Recommended system: ÄKTApurifier Recommended system: ÄKTAexplorer
w
Flo
F
st
rD
Fa
pe
XL
e™
Hy
Q
Q
e
15
30
os
os
Sample
ic
ar
ar
CE
CE
ra
ph
ph
1 mM EDTA,
UR
UR
Ce
250
Se
Se
ity
SO
SO
tiv
Q
Absorbance 260 nm
uc
nd
Co
pH 8.0 100
pH
50
1.0
230 nm
2000 254 nm
30.0
280 nm 80
6
0.8
0 100 200 ml
0.2 mM PMSF, pH 7.5 0.0 20.0 40.0 60.0 ml
gradient
7.5 15.0 min (mAU) mAU 280 nm 1500
1000 4
fusion 40 1000
20
20 20.0 500
20 300 60 200
0 0 60.0
Inject 0
200
60 ml 0.0 20
50 20.0
Polishing
UV 280 nm 2.0 4.0 6.0 8.0 Time (min)
A 230nm mAU
230 nm
%B BufferPrep.
mAU 254 nm 2000
pH 9.0
30.0 280 nm 80
600
• HiLoad 1000
40 400
0.1
200 10.0 40 • HiPrep 500
20 0
0 20 40 60 80 100 ml
• RESOURCE
0.0 20 0 Waste F8 Waste 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
Inject 20 40 60 80 100 120 140 160 ml
0 0
0 45 65 min 0.0 20.0 40.0 60.0 ml
w
Flo
F
st
rD
Fa
pe
XL
e™
Hy
Q
Q
e
15
30
os
os
Sample
ic
ar
ar
CE
CE
ra
ph
ph
1 mM EDTA,
UR
UR
Ce
250
Se
Se
ity
SO
SO
tiv
Q
Absorbance 260 nm
uc
nd
Co
pH 8.0 100
pH
50
1.0
230 nm
2000 254 nm
30.0
280 nm 80
6
0.8
0 100 200 ml
0.2 mM PMSF, pH 7.5 0.0 20.0 40.0 60.0 ml
gradient
7.5 15.0 min (mAU) mAU 280 nm 1500
1000 4
fusion 40 1000
20
20 20.0 500
20 300 60 200
0 0 60.0
Inject 0
200
60 ml 0.0 20
50 20.0
Polishing
UV 280 nm 2.0 4.0 6.0 8.0 Time (min)
A 230nm mAU
230 nm
%B BufferPrep.
mAU 254 nm 2000
pH 9.0
30.0 280 nm 80
600
• HiLoad 1000
40 400
0.1
200 10.0 40 • HiPrep 500
20 0
0 20 40 60 80 100 ml
• RESOURCE
0.0 20 0 Waste F8 Waste 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
Inject 20 40 60 80 100 120 140 160 ml
0 0
0 45 65 min 0.0 20.0 40.0 60.0 ml
w
Flo
F
st
rD
Fa
pe
XL
e™
Hy
Q
Q
e
15
30
os
os
Sample
ic
ar
ar
CE
CE
ra
ph
ph
1 mM EDTA,
UR
UR
Ce
250
Se
Se
ity
SO
SO
tiv
Q
Absorbance 260 nm
uc
nd
Co
pH 8.0 100
pH
50
1.0
230 nm
2000 254 nm
30.0
280 nm 80
6
0.8
0 100 200 ml
0.2 mM PMSF, pH 7.5 0.0 20.0 40.0 60.0 ml
gradient
7.5 15.0 min (mAU) mAU 280 nm 1500
1000 4
fusion 40 1000
20
20 20.0 500
20 300 60 200
0 0 60.0
Inject 0
200
60 ml 0.0 20
50 20.0
Polishing
UV 280 nm 2.0 4.0 6.0 8.0 Time (min)
A 230nm mAU
230 nm
%B BufferPrep.
mAU 254 nm 2000
pH 9.0
30.0 280 nm 80
600
• HiLoad 1000
40 400
0.1
200 10.0 40 • HiPrep 500
20 0
0 20 40 60 80 100 ml
• RESOURCE
0.0 20 0 Waste F8 Waste 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
Inject 20 40 60 80 100 120 140 160 ml
0 0
0 45 65 min 0.0 20.0 40.0 60.0 ml

valve Pump
Pump
Sample Pump
Air Sensor
concentration.
Buffer A
Mixer
Buffer A
Buffer B
Injection Valve
Pump NO
Buffer B NC
Gradient Mixer
formation
valve
(pH optional)
UV and Cond,
(pH optional)
Diversion to waste
Fractionation
Diversion to waste
Way of working
Explorer
!
Purifier
!
FPLC
!
Prime
!
Buffer Prep
! ! ! !
to increase purity
!
!
!
!
!
!
! Dedicated
configurations for
!
!
!
!
!
soon to come.....
the throughput.

of a button.

pump
A/D
PrimeView
converter
Air
sensors
pH
Flexibility
flowpath
Column scouting
Method wizard.
Reversed flow
Buffer selection

of a button.

pump
A/D
PrimeView
converter
Air
sensors
pH
Flexibility
flowpath
Column scouting
Method wizard.
Reversed flow
Buffer selection
Tel: +852 2811 8693 Tel: 0169 35 67 00 Tel: 21 417 7035
Australasia
Tel: +61 2 9899 0999
Fax: +61 2 9899 7511
Germany
Tel: 0761 4903 401
Fax: 0761 4903 405
Tel: +7 (095) 232 0250, 956 1137
Fax: +7 (095) 230 6377
ÄKTAdesign
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Tel: +358 (0)9 512 3940 Fax: +1 877 295 8102

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ÄKTAdesign FOR more information ABOUT One platform

THE COMPLETE ÄKTAdesign RANGE,

No matter whether the task is a quick sample

Simple Flexibility Sample

BufferPrep A B Autosampler A-900

BufferPrep A2 A1 C D ÄKTAFPLC builds on the world-wide success of

BufferPrep A B Autosampler A-900

BufferPrep A2 A1 C D ÄKTAFPLC builds on the world-wide success of

BufferPrep A B Autosampler A-900

BufferPrep A2 A1 C D ÄKTAFPLC builds on the world-wide success of

Inject 20 40 60 80 100 120 140 160 ml

Inject 20 40 60 80 100 120 140 160 ml

Inject 20 40 60 80 100 120 140 160 ml

Inject 20 40 60 80 100 120 140 160 ml

BufferPrep A B Autosampler A-900

BufferPrep A2 A1 C D ÄKTAFPLC builds on the world-wide success of

No matter whether the task is a quick sample

Simple Flexibility Sample

No matter whether the task is a quick sample

Simple Flexibility Sample

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