biocad 700e workstation for perfusion chromatography

BioCAD® 700E Workstation for Perfusion Chromatography®

Version 3 Series Software

Getting Started Guide

© Copyright 2000, 2011 Applied Biosystems

All rights reserved

For Research Use Only. Not for use in diagnostic procedures.

Information in this document is subject to change without notice. Applied Biosystems assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and accurate at the time of publication. In no event shall Applied Biosystems be liable for incidental, special, multiple, or consequential damages in connection with or arising from the use of this document.

Applied Biosystems, Perfusion Chromatography, POROS, the fractal icon, BioCAD, SCOUT, and RPM are registered trademarks of Applied Biosystems or its subsidiaries in the U.S. and certain other countries.

AB (Design), ABI, Applera, ChromatoCAD, ID, ImmunoDetection, SPRINT, Supervisor, and VISION are trademarks of Applied Biosystems or its subsidiaries in the U.S. and certain other countries.

Advantec is a registered trademark of Toyo Roshi International, Inc.

Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GMBH.

EZ is a trademark of Scientific Software, Inc.

Gilson is a registered trademark of Gilson, Inc.

Kel F is a registered trademark of 3M Company.

Kynar is a registered trademark of Union Carbide.

Microsoft and Windows are registered trademarks of Microsoft Corporation.

Pentium is a registered trademark of Intel Corporation.

RheFlex is a registered trademark and Rheodyne is a trademark of Rheodyne, L.P.

SLO-BLO is a trademark of Littlefuse, Inc.

SpectraSYSTEM and PushLoop are registered trademarks of Thermo Separation Products.

Swagelok is a registered trademark of Crawford Fitting Co.

Teflon and Tefzel are registered trademarks of E.I. Du Pont de Nemours and Co.

Tygon is a registered trademark of Norton Company.

Zip is a trademark and Iomega is a registered trademark of Iomega Corporation.

All other trademarks are the sole property of their respective owners.

Printed in the USA, 04/2011

Part Number 602478-01 Rev. B

WARNINGWARNINGBefore handling any chemicals, refer to the Material Safety Data Sheet provided by the manufacturer, and observe all relevant precautions.

AVERTISSEMENTAvant de manipuler des produits chimiques, veuillez consulter la fiche de sécurité du matériel fournie par le fabricant, et observer les mesures de précaution qui s’imposent.

Table of Contents

Table of Contents

Chapter 1 Before You Begin ............................................................1-1

Chapter 2 Starting Up and Connecting ....................................2-1

Chapter 3 Configuring, Plumbing, and Priming the Workstation ....................................................................... 3-1

3.1 Configuring ........................................................................... 3-2

3.2 Plumbing .............................................................................. 3-8

3.3 Priming................................................................................. 3-9

Chapter 4 Creating and Running a Method ...........................4-1

4.1 Creating a Method .................................................................. 4-1

4.2 Running the Method ................................................................ 4-9

Chapter 5 Running Templates ........................................................5-1

Chapter 6 Shutting Down ..................................................................6-1

Index

BioCAD ® 700E Workstation Getting Started Guide v

Table of Contents

vi Applied Biosystems

1Chapter

1 Before You Begin

Running your firstexperiment

The BioCAD® 700E Workstation Getting Started Guide is designed to help you quickly learn how to use the BioCAD 700E Workstation for Perfusion Chromatography®.

This guide contains brief procedures. For more detailed procedures and reference information, refer to the BioCAD 700E Workstation User’s Guide.

In the following chapters, you will:

• Start up and connect• Configure and prime the workstation• Create and run a method• Run a template• Shut down

NOTE: This guide assumes that your BioCAD 700E Workstation has been properly installed by a Applied Biosystems technical representative, that the workstation is plumbed in Tandem Column configuration, and that the 0.5 ml syringe is installed.

A quickintroductionto Windows

If you are unfamiliar with the Microsoft® Windows® environment, take a minute to read Section 1.6, A Quick Introduction to Microsoft Windows, in the BioCAD 700E Workstation User’s Guide.

BioCAD ® 700E Workstation Getting Started Guide 1-1

Chapter 1 Before You Begin

1
What you need To run the experiment in this guide, you need:
• Standard protein mix containing myoglobin, alpha chymotrypsinogen A, cytochrome C, and lysozyme

• MES, HEPES, Sodium acetate, HCl, NaOH, NaCl, and HPLC-grade water for buffer preparation

• POROS® HS M-series 4.6 mmD/100 mmL strong cation exchange column

Conditions The experiment shown in this guide is run using the following conditions:

Sample Standard protein mix

Buffer 20 mM cation buffers, pH range 4.5 to 7.5

Eluent Gradient

10 column volume gradient from 0 to 1,000 mM NaCl

Column POROS® HS M-series 4.6 mmD/100 mmL strong cation exchange column

Plumbing Tandem Column

Flow Rate 10 ml/min

Injection Mode

Autoloader

Syringe 0.5 ml

Sample Loop 100 µl

1-2 Applied Biosystems

1
Preparation Prepare the following and filter through a 0.22 or 0.45 µm filter
before use:

Sample 1 ml of a 5 mg/ml solution of myoglobin, alpha chymotrypsinogen A, cytochrome C, and lysozyme

Buffers 1. pH 4.5 buffer (500 ml)

33.3 mM MES + 33.3 mM HEPES + 33.3 mM Na acetate (adjust to pH 4.5 with HCl)

2. pH 7.5 buffer (500 ml)

33.3 mM MES + 33.3 mM HEPES + 33.3 mM Na acetate (adjust to pH 7.5 with NaOH)

3. HPLC-grade water (1 liter)

4. 3.0 M NaCl (500 ml)

For more information, see Section 3.2.2, Preparing pH Blend Mode Buffers, in the BioCAD 700E Workstation User’s Guide.


Chapter 1 Before You Begin

1


2Chapter

2 Starting Upand Connecting

In this chapter, you will:

• Power up the workstation• Start the BioCAD software• Connect buffers• Sparge buffers• Connect the sample

Powering upthe workstation

Power up the BioCAD 700E Workstation and external components in the following order:

• Turn on the compressed dry gas source (air or nitrogen) and regulate to 80 to 100 psi

• Turn on the helium source and regulate to 5 psi

• Power up the monitor and printer

• Power up the BioCAD 700E Workstation by turning on the power switch, located on the right side of the workstation, near the top of the panel


Chapter 2 Starting Up and Connecting

2

Starting theBioCAD

software

NOTE: If nothing appears on the monitor screen, check that the monitor is powered on, that the cable is connected between the monitor and the workstation, and adjust the contrast of the monitor display.

From Windows desktop, double-click the BioCAD icon to start the BioCAD software and display the Control Panel (Figure 2-1).

Figure 2-1 BioCAD Control Panel


Sparging buffers

2

Connectingbuffers

Place the buffer/solvent lines in the following solutions:

Sparging buffers Sparge the buffers/solvents to prevent outgassing and ensure pump flow rate precision and gradient accuracy:

1. Insert a buffer/solvent line and the corresponding sparge line through a bottle cap insert and into the sparge element (provided in the startup kit).

2. Submerge the sparge element and attached lines in the appropriate buffer/solvent or filtered deionized water.

3. Screw the bottle cap onto the bottle. Make sure the cap insert is vented.

4. Place the buffer/solvent reservoirs to the right of the workstation.

5. On the right side of the workstation, set the Helium On/Off switch to the On position.

6. Open the sparge valves for the reservoirs in use by turning the gas flow control knobs on the right side of the workstation. Open the valves until you see a steady stream of bubbles in the reservoir.

NOTE: For more information on sparging buffers, see Section 2.3.3, Connecting the Buffer/Solvent and Sparge Lines, in the BioCAD 700E Workstation User’s Guide.

Buffer/Solvent Line Solution

A pH 4.5 Buffer

B pH 7.5 Buffer

C Water

D 3.0 M NaCl

E and F Filtered deionized water

NOTE: Do not leave these lines dry.


Chapter 2 Starting Up and Connecting

2

Connectingthe sample

In this experiment, you will make an autoloader injection.

To connect the sample for an autoloader injection:

1. Place the sample in a small vessel and place the vessel in the sample compartment (Figure 2-2).

2. Insert sample line 1 in the sample vessel.

Figure 2-2 Preparing Sample for an Autoloader Injection

PB100519

Samplecompartment


3Chapter
3 Configuring,
Plumbing, andPriming theWorkstation


• Configure buffers, Autoloader, column, and data collection

• Plumb columns, tubing, and sample loop

• Prime buffer/solvent lines and sample line


Chapter 3 Configuring, Plumbing, and Priming the Workstation

3

3.1 Configuring

Configurationdefinition

A configuration specifies the physical setup of the BioCAD 700E Workstation, including buffers/solvents, samples, columns, and optional external devices.

DisplayingConfiguration

From the Control Panel, select Edit Config from the Config menu. The Configuration dialog box is displayed (Figure 3-1).

Figure 3-1 Configuration Dialog Box


Configuring

3

Configuringbuffers

To configure buffers:

1. Double-click buffer/solvent channel A in the Solvent Pump section of the Configuration dialog box.

Figure 3-2 Solvent Pump Section of Configuration Dialog Box

The Buffer/Solvent Library is displayed.

2. Select Cation Buffer - pH 4.5. Click OK.

3. Double-click buffer/solvent channel B to display the Buffer/Solvent Library.

4. Select Cation Buffer - pH 7.5. Click OK.

5. Double-click buffer/solvent channel C to display the Buffer/Solvent Library.

6. Select H20. Click OK.

7. Double-click buffer/solvent channel D to display the Buffer/Solvent Library.

8. Select NaCl + 3000 mM NaCl. Click OK.

Buffer/Solventchannels



3

Configuring theAutoloader

To configure the Autoloader for an autoloader injection:

1. In the Sample section of the Configuration dialog box (Figure 3-3), click the Configuration button under Autoloader.

Figure 3-3 Autoloader Configuration

The Autoloader dialog box is displayed (Figure 3-4).

Figure 3-4 Autoloader Dialog Box

Syringe 2. Select 0.5 ml from the Syringe Size drop-down list box.

Sample loop 3. Type 100 µl for the sample loop size.

Clickhere


Configuring

3

NOTE: Make sure the correct syringe and loop are installed. If not, refer to Section 3.1.4, Changing the Syringe and Sample Loop, in the BioCAD 700E Workstation User’s Guide.

4. Double-click sample channel 1 to display the Sample Library.

5. Click the New button to display the Sample Editor (Figure 3-5).

Figure 3-5 Sample Editor

6. Type Protein Standard in the Component field and in the Short Name text box. Click OK, then click OK again.

Protein Standard is displayed in sample channel 1.



3

Configuringcolumns

To configure the column plumbing option:

1. In the Plumbing section of the Configuration dialog box, select Tandem Columns from the Column Option drop-down list box (Figure 3-6).

NOTE: You will connect the column in Section 3.2, Plumbing. You will use a single column in this configuration with a union in place of column 2.

Figure 3-6 Plumbing Section of ConfigurationDialog Box

2. Double-click the top column position to display the Column Library.

3. Click the New button to display the Column Editor.

4. Type the part number and serial number (from the column label). Because you are using a POROS column, when you enter the column part number, all information other than the serial number is automatically filled in.

5. Click OK, then click OK again to return to the Configuration dialog box.

6. Do not configure the second column.

7. Type 0.020 inches for the Tubing ID.

8. Select the Flow Cell (Analytical or Semi-prep) installed on your system.

9. Click OK when configuration is complete.

Columnposition


Configuring

3

Configuringdata collection

To configure the detector channels to use for data collection:

1. In the Recorder section of the Configuration dialog box (Figure 3-7), click Setup.

Figure 3-7 Recorder Section of Configuration Dialog Box

The Data Recorder Configuration dialog box is displayed (Figure 3-8).

Figure 3-8 Data Recorder Configuration Dialog Box

2. Select the UV/Vis #1, pH, Conductivity, and Pressure check boxes to enable the collection of this data in the electronic Strip Chart Recorder and the data file.

NOTE: Because you will look at only one UV/Vis wavelength, you will not enable UV/Vis #2 at this time.

3. Type 1 for the Buffer Duration to set the duration of the electronic Strip Chart Recorder.

4. Click OK to return to the Configuration dialog box.

Click here



3

3.2 Plumbing

In this experiment, you will leave the plumbing in Tandem Column configuration. However, you will run the experiment using one column with a union in place of column 2.

To plumb the columns, tubing, and sample loop:

1. Selecting Show Column Plumbing from the Config menu in the Control Panel.

The Tandem Columns plumbing diagram is displayed.

NOTE: If the plumbing diagram is for a plumbing configuration other than Tandem Columns, configure Tandem Columns in the Configuration dialog box. See “Configuring columns” on page 3-6.

2. Verify that your plumbing is connected properly. Verify that the injection and column switching valves are plumbed with orange 0.020-inch ID tubing.

3. Connect the POROS® HS M-series 4.6 mmD/100 mmL column in the Column 1 position.

4. Connect a union in the Column 2 position.

5. Install a 100 µl sample loop on the injection valve.

For help on plumbing columns, tubing, and the sample loop, refer to Section 3.1, Plumbing the Workstation, in the BioCAD 700E Workstation User’s Guide.


Priming

3

3.3 Priming

After configuring and plumbing the workstation, prime the buffer/solvent lines and the sample line.

Primingbuffer/solvent

lines

Prime the buffer/solvent lines to fill the lines and remove air bubbles. Before priming, verify that each buffer/solvent line is submerged in the appropriate buffer/solvent. For more information, see “Connecting buffers” on page 2-3.

To prime the buffer/solvent lines:

1. Connect the 60 ml syringe (provided in the startup kit) to the Luer-Lok® fitting at the end of the vent tube connected to the prime/purge valve (Figure 3-9). Open the valve one-half to one full turn to the left (counterclockwise).

Figure 3-9 Syringe Attached to Luer-Lok Fitting on Prime/Purge Valve

2. Select Purge System from the Control Panel Blend menu.

Prime/purgevalve

Vent tube



3

The Purge Segment Setup dialog box is displayed (Figure 3-10).

Figure 3-10 Purge Segment Setup Dialog Box

3. Select buffer/solvent channel A and set it for 100%.

4. Type a flow rate of at least 20 ml/min.

5. Type 30 ml in the volume text box.

6. Select Bypass for the Flow Direction at End of Purge.

7. Click OK to start the purge segment.

8. Prime the line by pulling 20 ml of buffer/solvent into the syringe you attached to the prime/purge valve.

9. Close the prime/purge valve by turning it clockwise and remove the syringe from the vent tube.

10. Stop the pump by clicking the Stop button on the Control Panel or by pressing the Pump button on the front panel of the workstation.

11. Repeat this procedure for channels B through F. Prime the four channels you will use (A through D) with the appropriate buffer/solvent. Prime unused lines (E and F) with filtered deionized water.


Priming

3

Primingsample line

To prime the sample line:

1. Place sample line 1 from the autoloader in the sample vessel.

2. Select sample line 1: Protein Standard from the Sample Loader drop-down list box in the Sample section of the Control Panel (Figure 3-11).

Figure 3-11 Sample Section of Control Panel

3. Select Prime from the Sample menu in the Control Panel.

The syringe moves up and down to draw sample through sample line 1 to the injection valve.



3


4Chapter

4 Creating andRunning aMethod


• Create a method• Run the method

4.1 Creating a Method

To create a method, you will assemble the following parts in the Method Editor:

• General Settings—To set flow rate, flow direction, wavelength, and pH/conductivity monitors

• Equilibrate block—To purge the system and equilibrate the column

• Load block—To inject the sample onto the column

• Wash block—To wash the column

• Elute block—To elute the sample from the column

• Method events—To zero the detector


Chapter 4 Creating and Running a Method

4

Displaying theMethod Editor

To display and set up the Method Editor:

1. Select Method Editor from the Window menu.

The Method Editor is displayed (Figure 4-1).

Figure 4-1 Method Editor

2. Click the EQ, L, W, and EL buttons at the top of the window to add Equilibrate, Load, Wash, and Elute blocks to the method.


Creating a Method

4

General Settings To set the General Settings:

1. Double-click the General Settings line.

The General Settings dialog box is displayed (Figure 4-2).

Figure 4-2 General Settings Dialog Box

2. Type 10 in the Flow Rate text box and select ml/min from the units drop-down list box.

3. Type 280 in the UV Wavelength text box.

4. Click Column 1 to place column 1 inline.

5. Click Inline to place the pH and conductivity monitors inline.

6. Click OK.



4

Equil block To add an Equil block:

1. Select the Equil block line.

Segment buttons (S, G, and P) are now active.

2. Click the P segment button to add a Purge segment.

The Purge Segment Setup dialog box is displayed (Figure 4-3).

Figure 4-3 Purge Segment Setup Dialog Box

3. Click the Setup button, and select pH instead of Percentage for the blend mode. Click OK.

4. Enter initial blend conditions. Type 20 mM for buffer concentration, pH 4.5, and 0 mM for initial eluent (NaCl) concentration.

5. Click Column 1 to place the column inline after the purge.

6. Click OK to add the Purge segment.

NOTE: When you run the method, the Purge segment rapidly equilibrates the system by purging at a high flow rate with the column offline. It then places the column inline.


Creating a Method

4

7. Click the S segment button to add a Step segment.

NOTE: If segment buttons are dimmed, click the blank line below the Purge segment.

The Step Segment Setup dialog box is displayed (Figure 4-4) with the initial blend conditions you entered in the previous Purge segment.

Figure 4-4 Step Segment Setup Dialog Box

8. Type 10 for duration and select CV from the units drop-down list box.

NOTE: If CV units are not available, make sure you configured a column and placed the column inline at the end of the Purge segment.

Hint: You can change the Segment Name to Equilibrate Column. If you do not type a segment name, it is labeled Step Segment instead of Equilibrate Column in the method.

9. Click OK to add the Step segment.

NOTE: When you run the method, the Step segment equilibrates the column for 10 column volumes.



4

Load block To add a Load block:

1. Select the Load block line.

2. Click the I segment button to add an Inject segment.

The Sample Loader Inject Segment dialog box is displayed (Figure 4-5).

Figure 4-5 Sample Loader Inject Segment Dialog Box

3. Select sample channel 1: Protein Standard.

4. Type 20 for the Injection Amount and select µl from the units drop-down list box.

5. Click OK to add the Inject segment.

Wash block To add a Wash block:

1. Select the Wash block line.


The Step Segment Setup dialog box is displayed (Figure 4-4) with the same blend conditions set in the previous Step segment (Equil block).

3. Type 2 for the wash duration and select CV from the units drop-down list box.



Creating a Method

4

Elute block To add an Elute block:

1. Select the Elute block line.

2. Click the G segment button to add a Gradient segment.

The Gradient Segment Setup dialog box displays the same blend conditions that were set in the previous Step segment (Wash block) (Figure 4-6).

Figure 4-6 Gradient Segment Setup Dialog Box

3. Type 1000 mM for the final eluent concentration.

NOTE: Because you entered the NaCl concentration in the Configuration dialog box, the BioCAD 700E Workstation will blend the correct gradient.

4. Type 10 for the gradient duration and select CV from the units drop-down list box.

5. Click OK to add the Gradient segment.


The Step Segment Setup dialog box is displayed (Figure 4-4) with the final blend conditions you entered in the previous Gradient segment.



4

7. Type 5 for the duration to hold the gradient and select CV from the units drop-down list box.

NOTE: When a method runs, the Step segment holds the final gradient blend conditions for the duration you specify.

8. Type Hold Gradient in the Segment Name text box.


Addingmethod events

Method events are additional functions you can add to customize the method.

NOTE: Method events are not displayed in Block view (the view currently displayed). To display the method in more detail before adding events, select Sequence View from the Display menu.

To add method events:

1. Select the Equilibrate Column Step segment in the Equil block.

2. Click the Add Event button.

The Add Event dialog box is displayed.

3. Scroll down to the Detector Events. Select the Zero UV Detector event and click Add.

The Zero UV Detector dialog box is displayed.

4. Enter 10 CV (the volume that corresponds to the end of the equilibrate segment). Click OK.

NOTE: When you run the method, this event zeroes the detector after the column is equilibrated, but before you make an injection.

5. Save the method by selecting Save As from the File menu. Type TESTMETH for the method name.

The method is saved as TESTMETH.MET in the C:\BIOCAD\METHODS directory.


Running the Method

4

4.2 Running the Method

Before running the method, you must tell the system where to store the data.

Setting datastorage options

To set data storage options:

1. Select Data Storage from the Options menu in the Method Editor window.

The Set Data File Name and Directory dialog box is displayed (Figure 4-7).

Figure 4-7 Set Data File Name and Directory Dialog Box

2. Type TESTDATA as the file name for the data.

3. Click the Auto File Extension check box. This function saves subsequent method runs with the same file name, but sequential file extensions (.B01, .B02, and so on).

4. Create a new directory in which to store the data by clicking the Make Dir button. Type TESTRUN as the directory name and click OK.

Hint: To easily find data files related to specific users and projects, create subdirectories in the C:\BIOCAD\DATA directory for different projects, users, or data sets.

5. Click OK.

6. To save these data storage options for future use, select Save from the File menu in the Method Editor window.



4

Running themethod

To run TESTMETH.MET, select Run Method from the File menu In the Method Editor window.

Viewing themethod run

After starting the method run, you can observe the status of the run in two windows:

• Strip Chart Recorder• Status window

Close the Method Editor window by double-clicking the Control Menu box (minus sign) in the top left corner of the window.

The Strip Chart Recorder is located below the Control Panel. The Status window is located to the right of the Control Panel (Figure 4-8).

Figure 4-8 Strip Chart Recorder and Status Window

NOTE: If either window is not displayed, select Strip Chart Recorder or Status from the Window menu.

If the run is acceptable, you can:

• Run a template (see Chapter 5, Running Templates)

• Analyze the data (see the Data Analysis User’s Guide)


Chapter5

5 Running Templates

In this chapter, you will create and run a pH Map template.

Templates A BioCAD template is a predefined program set up to perform a sequence of runs in which one variable changes from run to run. Templates help you systematically investigate the effects of varying a single parameter.

Templates andData Analysis

Template and Data Analysis features make systematic methods development easy and efficient.

Create method

Run method

Create template(Method Editor)

(Method Editor)

Run template

View chromatograms(Data Analysis)Select optimized

(Data Analysis)chromatogram

Open method(Data Analysis)


Chapter 5 Running Templates

5

Creating andrunning a

pH Map template

To create and run a pH Map template:

1. Display the Method Editor by selecting Method Editor from the Window menu.

2. Select Open from the File menu to open TESTMETH.MET, the method you created in Section 4.1, Creating a Method.

3. Select pH Map from the Templates menu.

The pH Map Template dialog box is displayed (Figure 5-1).

Figure 5-1 pH Map Template

4. Enter pH values of 5.0, 5.5, 6.0, 6.5, 7.0, and 7.5.

NOTE: Typically, when you run a pH Map template, you enter 0.5 pH unit increments. If you are familiar with the characteristics of the sample, or if you are investigating more specific pH effects, you can set up the template in smaller pH unit increments.

5. Click Run.

The method runs six times, once at each pH value.


pH Map template

5

NOTE: When you created the method, you set the data storage options to name the data file TESTDATA, using the auto file extension feature. The files collected using this template are named TESTDATA.B01, TESTDATA.B02, and so on. The template also creates a group file called TESTDATA.GRO.

6. Observe the running template in the Status window and the Strip Chart Recorder. For more information, “Viewing the method run” on page 4-10.

7. When the runs are complete, view the chromatograms in Data Analysis. For more information, see the Data Analysis User’s Guide.


Chapter 5 Running Templates

5


Chapter6

6 Shutting Down

Powering down To power down the BioCAD 700E Workstation when the method is complete:

CAUTIONFailure to leave the pH and conductivity monitors in a 1 M salt solution and to run the SYSCLEAN method after daily use may result in system downtime.

1. Click the Bypass option button in the Column section of the Control Panel to take the column out of line.

2. Purge the system with a 1 M salt solution with the pH and conductivity detectors inline. For more information, see Section 5.2.4, Purging the System, in the BioCAD 700E Workstation’s User’s Guide.

3. Run the SYSCLEAN method, supplied with the system:

• Place all buffer/solvent lines and sample lines in filtered deionized water.

• Select Open from the File menu in the Method Editor window to display the Open File dialog box.


Chapter 6 Shutting Down

6
• Select SYSCLEAN.MET from the C:\BIOCAD\METHODS directory and click OK.
You may see some configuration mismatch errors, which are usually due to differences in buffer/solvent and column configuration. Click Update Method with System Configuration to accept changes.

• Select Run Method from the File menu. This method takes the column, pH monitor, and conductivity monitor out of line.

4. When the SYSCLEAN method is complete, close the BioCAD software by selecting Exit BioCAD from the File menu in the Control Panel.

5. To exit Microsoft Windows, click Start, then click Shut Down.

6. Power down the components of the workstation in any order.

7. Close the sparge valves by turning the gas flow control knobs on the right side of the workstation.

8. Set the Helium On/Off switch to the Off position.

9. Turn off gas and helium sources.


IndexI

DEX

N

BBuffers

configuring 3-3description 1-2preparing 1-3priming 3-9sparging 2-3

CColumn

bypassing 6-1configuring 3-6connecting 3-8description 1-2part number 3-6placing inline 4-3, 4-4plumbing 3-8POROS 1-2serial number 3-6

Compressed gas pressure 2-1Conductivity monitor

placing inline 4-3shutting down 6-1

Configurationbuffers 3-3column 3-6data channels 3-7definition 3-2plumbing 3-6sample 3-4Sample Loader 3-4sample loop 3-4syringe 3-4

Configuration dialog box 3-2

DData channels, configuring 3-7Data files, setting directory 4-9Data storage, setting 4-9

EElute block 4-1, 4-7Equil block 4-1, 4-4Events, method 4-8Experiment

conditions 1-2requirements 1-2

FFlow rate 1-2, 4-3

GGas pressure 2-1General Settings 4-1, 4-3Gradient

conditions 1-2, 4-7holding 4-8

Gradient segment 4-7

HHelium pressure 2-1Hold, gradient 4-8

BioCAD® 700E Workstation Getting Started Guide Index-1

I

DEX

N
IInject segment 4-6Injection mode 1-2, 2-4, 4-6Injection volume 4-6
LLoad block 4-1, 4-6

MMethod

creating 4-1events 4-1, 4-8parts of 4-1running 4-9saving 4-8setting data storage 4-9template 5-1viewing status 4-10

Method Editor 4-2Microsoft Windows, quick

introduction 1-1

PpH map

buffer preparation 1-3creating template 5-2running 5-2

pH monitorplacing inline 4-3shutting down 6-1

Plumbingcolumns 3-8configuration 3-6connecting 3-8description 3-8sample loop 3-8Tandem Column 3-8tubing 3-8

POROS column 1-2Powerdown 6-1Powerup sequence 2-1, 2-3Preparing

buffers 1-3sample 1-3

Pressurecompressed gas 2-1helium 2-1

Primingbuffer/solvent lines 3-9sample line 3-11

Purging 4-4

RRequirements for experiment 1-2

SSample

connecting 2-4preparing 1-3priming 3-11

Sample loop size 1-2, 3-4Saving

data files 4-9method 4-8

Shutdown 6-1Software

shutting down 6-2

Index-2 Applied Biosystems

I

DEX

N

Spargingbuffers 2-3pressure 2-1rate 2-3

Standard used 1-2Startup sequence 2-1, 2-3Status of method 4-10Status window 4-10Step segment 4-5, 4-6, 4-7Strip Chart Recorder 4-10Syringe size 1-2, 3-4SYSCLEAN method 6-1

TTemplate

creating 5-2definition 5-1running 5-2

UUV/Vis detector, zeroing 4-8UV/Vis wavelength 4-3

VViewing method run 4-10

WWash block 4-1, 4-6Wavelength, UV/Vis 4-3Windows, quick introduction 1-1

ZZeroing UV/Vis detector 4-8

BioCAD® 700E Workstation Getting Started Guide Index-3

I

DEX

N

Index-4 Applied Biosystems

Headquarters850 Lincoln Centre DriveFoster City, CA 94404 USA

Phone: +1 650.638.5800Toll Free (In North America): +1 800.345.5224Fax: +1 650.638.5884

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Printed in the USA, 12/2000Part Number 602478-01 Rev. A

biocad 700e workstation for perfusion chromatography

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