ÄKTA system Training Guide · ÄKTAexplorer am ers ha m bio scie nce am er sham bio ci n a m e r s...
Transcript of ÄKTA system Training Guide · ÄKTAexplorer am ers ha m bio scie nce am er sham bio ci n a m e r s...
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ÄKTAexplorer
amersham bioscience
amersham bioscience
amersham bioscience
amersham bioscience
ÄKTA system Training GuideExplorer 100 / 10
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ÄKTAexplorer
amersham bioscience
amersham bioscience
amersham bioscience
amersham bioscienceBox
MonitorConductivitypH
UV Detector3 Wavelengths
Pumps
ColumnValves
Injector
SampleValve
Buffers
pH/cell
electrode
Sampleholder
Pump A
Buffer switchingvalve
OutletValve
ÄKTAexplorer
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23 4 5
6781
B
4
A
Systempumps
Mixer
FractioncollectorFrac-950
Conductivitymonitor
pH detector
In-linefilter
Samplepump
Buffer selectvalve (V6)
Injectionvalve (V1)
Reverseflow valve (V7)
Outletvalve (V4)
Sample selectionvalve (V5)
Columnselectionvalve (V2)
Columnselectionvalve (V3)
3 Wavelengthdetector
23 4 5
6781
23 4 5
6781
23 4 5
6781
23 4 5
6781
24
7
3
165
24
7
3
165
UV
Cond
pH
ÄKTAexplorer 100
Airsensor
23 4 5
6781
B
4
A
Systempumps
Mixer
FractioncollectorFrac-950
Conductivitymonitor
pH detector
In-linefilter
Samplepump
Buffer selectvalve (V6)
Injectionvalve (V1)
Reverseflow valve (V7)
Outletvalve (V4)
Sample selectionvalve (V5)
Columnselectionvalve (V2)
Columnselectionvalve (V3)
3 Wavelengthdetector
23 4 5
6781
23 4 5
6781
23 4 5
6781
23 4 5
6781
24
7
3
165
24
7
3
165
UV
Cond
pH
ÄKTAexplorer 100 Air
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23 4 5
6781
B
4
A
Systempumps
Mixer
FractioncollectorFrac-950
Conductivitymonitor
pH detector
In-linefilter
Buffer selectvalve (V6)
Injectionvalve (V1)
Outletvalve (V4)
Columnselectionvalve (V2)
Columnselectionvalve (V3)
3 Wavelengthdetector
23 4 5
6781
23 4 5
6781
23 4 5
6781
UV
Cond
pH
ÄKTAexplorer 10
23 4 5
6781
B
4
A
Systempumps
Mixer
FractioncollectorFrac-950
Conductivitymonitor
pH detector
In-linefilter
Samplepump
Buffer selectvalve (V6)
Injectionvalve (V1)
Outletvalve (V4)
Sample selectionvalve (V5)
Columnselectionvalve (V2)
Columnselectionvalve (V3)
3 Wavelengthdetector
23 4 5
6781
23 4 5
6781
23 4 5
6781
23 4 5
6781
UV
Cond
pH
ÄKTAexplorer 10 S
24
7
3
165
24
7
3
165
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ÄKTAexplorer 10 XT
23 4 5
6781
B
4
A
Systempumps
Mixer
FractioncollectorFrac-950
Conductivitymonitor
pH detector
In-linefilter
Buffer selectvalve (V6)
Injectionvalve (V1)
Outletvalve (V4)
Columnselectionvalve (V2)
Columnselectionvalve (V3)
3 Wavelengthdetector
23 4 5
6781
23 4 5
6781
23 4 5
6781
UV
Cond
pH
24
7
3
165
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PUMP P-900
Buffer Acid /Alkali WaterSalt
Proportion Valve
Pump A
Mixer
A2 A1 B2 B1
Pump B
GRADIENT MIXERÄ KTA 10 mls/min <6 mls/min
6 -14 mls/min0.6 ml mixer2 ml mixer
Ä KTA 100 mls/min <30 mls/min30 - 50 mls/min50 - 100 mls/min
2 mls mixer5 mls mixer12 mls mixer
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PUMP P-900 Inlets
Water Salt
Buffer Select Valve IV-908
Pump A Pump B
Mixer
A2 A1B1 B2
SwitchingValves
12
34 5
67
8
A11A12
A13A14
A15A16
A17A18
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PUMP P-900 Manually Priming Buffer Inlet Lines
Open
Draw Air out ofBuffer Line
Manual Flowpath
A1 or A2 BufferValveA
Manual Flowpath
B1 or B2 BufferValveB
Pump A Pump B
Common inlet A2 A1 B2 B1Common inlet
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PUMP P-900 Back - Wash Lines
20% Ethanol : Water
Pump Back-WashLines
Wash OUT
WashIN
BufferIN
BufferOUT
High pressurePiston seal
Ball outletcheck valve
Ball inletcheck valve
Piston
Backwash pump diaphragm
IN-LINE one-wayvalve
Syringe fitting
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LOAD (sample) INJECT (sample) LOAD (wash loop)
1
7
6
5
2
3
4
Pump
Column
Waste
PV-9081
2
34 5
6
7
8
Sample
1
7
6
5
2
3
4
Pump
Column
Waste
PV-9081
2
34 5
6
7
8
Sample
1
7
6
5
2
3
4
Pump
Column
Waste
PV-9081
2
34 5
6
7
8
Sample
ON-LINE FILTERS
P-901 ( 100 ml/min Pump )High Flow Fibre Filter
Polymer Support
P-903 ( 10 ml/min Pump )Low Flow HPLC Filter
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1
7
6
5
2
3
4
PUMPLOADSAMPLE
Waste
COLUMN
Waste
LOAD
1
7
6
5
2
3
4
PUMP
Waste
COLUMN
Waste
INJECT
1
7
6
5
2
3
4
PUMP
Waste
COLUMN
Waste
WASTE
INJECTION VALVE INV- 907
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SAMPLE LOADING METHOD
1
7
6
5
2
3
4
PUMPLOADSAMPLE
Waste
COLUMN
Waste Superloop
Luer LockAdapter
1
7
6
5
2
3
4
PUMPLOADSAMPLE
Waste
COLUMN
Waste
Needle Port
Always tighten by hand with needle inserted
Do Not Overtighten
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Volume Max. allowed column pressure
Code No.
10 ml 4 Mpa 18-1113-8150 ml 4 Mpa 18-1113-82150 ml 2 Mpa 18-1023-85
SUPERLOOPS
10 ml
150 ml
50 ml
BUFFERFLUSH
SAMPLELOAD
SAMPLEINJECT
SAMPLEFLUSH
WASTEPUMP orSYRINGE
COLUMN COLUMN
Plunger
Buffer
Sample
EtchedGroove
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Needle
NeedleAdapter
INV-907 Injection Valve - Injection Port
PEEK Valve Body
Valve Seal
Injection Needle Port
NeedleTube
Ferrule
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INV-907 Injection Valve - Luer Adapter
PEEK Injection Valve BodyPEEK Injection Valve Body
PEEK Injection Luer AdaptorPEEK Injection Luer Adaptor
Valve SealValve Seal
Valve Channel Plate Valve Channel Plate
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Flow
End Beginning
PEEK Tubing High Resistanceto Flow
Low Resistanceto Flow
Fluid Dynamics
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Flow - Injecting a Loop
Fluid DynamicsGeneral
The way we imagine it Flows
The way it Actually Flows
Bubble Segmentation
• Partial Loading Loop = No more than 1/2 Loop Volume• Total Loading Loop = Inject 2X Loop Volume
50ul Load
60ul Load
100ul Load
200ul Load
Waste
100ul Loop
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LOADING A SAMPLE LOOP
1
7
6
5
2
3
4
Sample
Pump
Column
Inject
Load
SamplepumpP-950
Injection valveINV-907
Waste
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LOADING A SUPER LOOP
1
7
6
5
2
3
4
Sample
Pump
Column
Inject
Load
Waste
Sample
Buffer
Injection valveINV-907
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MULTI-LOADING A SAMPLE LOOP
1
7
6
5
2
3
4
Pump
Inject
Load
Column
PV-908
Samples
1
2
3 4 56
7
8
I/O
Injection valveINV-907
WasteWash
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MULTI-LOADING A SUPER LOOP
Waste
1
7
6
5
2
3
4
PumpColumn
Inject
Load
PV-9081
2
3 4 56
7
8
I/O
Wash
Samples
S3 S4 S5 S6 S8
Injection valveINV-907
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DIRECT LOADING ON TO COLUMN
PumpColumn
Extra Tube
Inject
Load
1/16” or 1/8”Tubing
Sample
1
7
6
5
2
3
4
Waste
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REVERSE FLOW VALVE ( INV-907 )
Column
Pump
Downward FlowPosition 1
Extra Tube
MONITOR
StopPlug
17
65
2
34
1
Column Extra Tube
MONITOR
StopPlug
2
Upward FlowPosition 2
Pump7
65
34
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OUTLET VALVE PV-908
Default
To Fraction Collector
DETECTOR
F1
F2
F3F4
F5F6
F7
F8
I/O
Waste
F3 F5 F6
F7
F8F4
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0
%B 10
0
50
Elu
ate
Fra
cti
on
atio
nP
eak
Fra
ctio
na
tio
nF
low
thro
ug
hF
ract
ion
ati
on
0%
B10
0%
B
Start
End
A1
A2
A3
……
……
……
….
Injectm
AU
0
500
250
Waste
Fra
ctio
n C
olle
ctio
n
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HiTrap Connections
M6 Male
1/16” Female
1/16” Male
1/16” Female
M6 Male
M6 Female
1/16” Male
M6 male / 1/16” Female
18-1112-57
18-1112-55
Flow Cell Inlet
HiTrap Column
18-1112-58
M6 female / 1/16” male
1/16” Fingertight Fitting
Pheny l Sepharos e
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Detector
Multiple HiTrap Arrangment ( Series )
Phenyl S
epharose
Detector
Phenyl S
epharoseP
henyl Sepharose
Detector
Phenyl S
epharoseP
henyl Sepharose
Phenyl S
epharose