ÄKTA system Training Guide

Transcription

1 ÄKTA system Training Guide Explorer 00 / 0 amersham bioscience amersham bioscience amersham bioscience amersham bioscience ÄKTAexplorer

2 ÄKTAexplorer Buffers Injector Box amersham bioscience Monitor Conductivity ph amersham bioscience Valves UV Detector Wavelengths amersham bioscience Outlet Valve Pumps amersham bioscience ÄKTAexplorer Pump A Buffer switching valve Valve holder ph/cell electrode

3 ÄKTAexplorer 00 System pumps A Mixer In-line filter B Injection valve (V) Reverse flow valve (V) selection valve (V) UV Cond ph Wavelength detector Conductivity monitor ph detector pump Fraction collector Frac-90 Buffer select valve (V) Outlet valve (V) selection valve (V) selection valve (V) ÄKTAexplorer 00 Air System pumps A Mixer In-line filter B Injection valve (V) Reverse flow valve (V) selection valve (V) UV Cond ph Wavelength detector Conductivity monitor ph detector Air sensor pump Fraction collector Frac-90 Buffer select valve (V) Outlet valve (V) selection valve (V) selection valve (V)

4 ÄKTAexplorer 0 System pumps A Mixer In-line filter B Injection valve (V) selection valve (V) UV Cond ph Wavelength detector Conductivity monitor ph detector Fraction collector Frac-90 Buffer select valve (V) Outlet valve (V) selection valve (V) ÄKTAexplorer 0 S System pumps A Mixer In-line filter B Injection valve (V) selection valve (V) UV Cond ph Wavelength detector Conductivity monitor ph detector pump Fraction collector Frac-90 Buffer select valve (V) Outlet valve (V) selection valve (V) selection valve (V)

5 ÄKTAexplorer 0 XT System pumps A Mixer In-line filter B Injection valve (V) selection valve (V) UV Cond ph Wavelength detector Conductivity monitor ph detector Fraction collector Frac-90 Buffer select valve (V) Outlet valve (V) selection valve (V)

6 PUMP P-900 Mixer Pump A Pump B Proportion Valve A A B B Acid /Alkali Buffer Salt Water Ä KTA 0 mls/min Ä KTA 00 mls/min GRADIENT < mls/min - mls/min <0 mls/min 0-0 mls/min 0-00 mls/min MIXER 0. ml mixer ml mixer mls mixer mls mixer mls mixer

7 PUMP P-900 Inlets Mixer Pump A Pump B Switching Valves Buffer Select Valve IV-908 A A B B 8 Water Salt A A A A A A A8 A

8 PUMP P-900 Manually Priming Buffer Inlet Lines Pump A Pump B Common inlet A A Common inlet B B Manual Flowpath Manual Flowpath A or A BufferValveA B or B BufferValveB Open Draw Air out of Buffer Line

9 PUMP P-900 Back - Wash Lines Pump Back-Wash Lines Syringe fitting Wash OUT IN-LINE one-way valve High pressure Piston seal Buffer OUT Ball outlet check valve 0% Ethanol : Water Piston Backwash pump diaphragm Wash IN Buffer IN Ball inlet check valve

10 ON-LINE FILTERS P-90 ( 0 ml/min Pump ) Low Flow HPLC Filter P-90 ( 00 ml/min Pump ) High Flow Fibre Filter Polymer Support LOAD (sample) INJECT (sample) LOAD (wash loop) Pump Pump Pump 8 PV PV PV-908

11 INJECTION VALVE INV- 90 COLUMN LOAD SAMPLE PUMP LOAD COLUMN PUMP INJECT COLUMN PUMP WASTE

12 SAMPLE LOADING METHOD COLUMN LOAD SAMPLE PUMP Needle Port Always tighten by hand with needle inserted Do Not Overtighten COLUMN LOAD SAMPLE PUMP Luer Lock Adapter Superloop

13 SUPERLOOPS Buffer 0 ml 0 ml 0 ml Plunger BUFFER FLUSH SAMPLE LOAD SAMPLE INJECT SAMPLE FLUSH Etched Groove WASTE PUMP or SYRINGE COLUMN COLUMN Volume Max. allowed column pressure Code No. 0 ml Mpa ml Mpa ml Mpa 8-0-8

14 INV-90 Injection Valve - Injection Port PEEK Valve Body Injection Needle Port Valve Seal Ferrule Needle NeedleTube Needle Adapter

15 INV-90 Injection Valve - Luer Adapter PEEK PEEK Injection Valve Valve Body Body PEEK PEEK Injection Luer Luer Adaptor Valve Valve Seal Seal Valve Valve Channel Plate Plate

16 Fluid Dynamics Flow High Resistance to Flow PEEK Tubing Low Resistance to Flow End Beginning

17 Fluid Dynamics Flow - Injecting a Loop General The way we imagine it Flows The way it Actually Flows Bubble Segmentation 0ul Load 00ul Loop 0ul Load 00ul Load 00ul Load Partial Loading Loop = No more than / Loop Volume Total Loading Loop = Inject X Loop Volume

18 LOADING A SAMPLE LOOP Load Inject Injection valve INV-90 Pump pump P-90

19 LOADING A SUPER LOOP Load Inject Injection valve INV-90 Pump Buffer

20 MULTI-LOADING A SAMPLE LOOP Load Inject Injection valve INV-90 Pump PV I/O s Wash

21 MULTI-LOADING A SUPER LOOP Injection valve INV-90 Pump Load Inject PV-908 I/O 8 s S S S S Wash S8

22 DIRECT LOADING ON TO COLUMN Pump Inject Load Extra Tube / or /8 Tubing

23 REVERSE FLOW VALVE ( INV-90 ) Pump Extra Tube Stop Plug MONITOR Downward Flow Position Pump Extra Tube Stop Plug MONITOR Upward Flow Position

24 OUTLET VALVE PV-908 DETECTOR Default To Fraction Collector F F F F8 I/O F F F F F F F F F F8

25 Flowthrough Fractionation Eluate Fractionation Peak Fractionation 0%B 00%B %B Start End Fraction Collection mau 00 Inject 0 0 A A A.

26 Phenyl Sepharose HiTrap Connections / Female / Male 8-- / Fingertight Fitting / Female M Male 8-- M male / / Female HiTrap M Male M Female 8--8 M female / / male / Male Flow Cell Inlet

27 Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose Multiple HiTrap Arrangment ( Series ) Detector Detector Detector