Thermo Fisher Scientific BioCAD® 700E Workstation Quick start guide

Brand: Thermo Fisher Scientific

Model: BioCAD® 700E Workstation

Category: Software manuals

Type: Quick start guide

BioCAD® 700E Workstation for

Perfusion Chromatography®

Version 3 Series Software

Getting Started Guide

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

WARNINGWARNING

Before handling any chemicals, refer to the Material Safety Data Sheet

provided by the manufacturer, and observe all relevant precautions.

AVERTISSEMENT

Avant 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

BioCAD® 700E Workstation Getting Started Guide v

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

Table of Contents

vi Applied Biosystems

Chapter

BioCAD® 700E Workstation Getting Started Guide 1-1

1 Before You Begin

Running your first

experiment 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 quick

introduction

to 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.

Chapter 1 Before You Begin

1-2 Applied Biosystems

1What 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

BioCAD® 700E Workstation Getting Started Guide 1-3

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

1-4 Applied Biosystems

Chapter

BioCAD® 700E Workstation Getting Started Guide 2-1

2 Starting Up

and Connecting

In this chapter, you will:

•Power up the workstation

•Start the BioCAD software

•Connect buffers

•Sparge buffers

•Connect the sample

Powering up

the 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

2-2 Applied Biosystems

Starting the

BioCAD

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

BioCAD® 700E Workstation Getting Started Guide 2-3

Connecting

buffers 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

2-4 Applied Biosystems

Connecting

the 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

Sample

compartment

Chapter

BioCAD® 700E Workstation Getting Started Guide 3-1

3 Configuring,

Plumbing, and

Priming the

Workstation

In this chapter, you will:

•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

3-2 Applied Biosystems

3.1 Configuring

Configuration

definition A configuration specifies the physical setup of the BioCAD 700E

Workstation, including buffers/solvents, samples, columns, and

optional external devices.

Displaying

Configuration 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

BioCAD® 700E Workstation Getting Started Guide 3-3

Configuring

buffers 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/Solvent

channels

Chapter 3 Configuring, Plumbing, and Priming the Workstation

3-4 Applied Biosystems

Configuring the

Autoloader 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.

Click

here

Configuring

BioCAD® 700E Workstation Getting Started Guide 3-5

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.

Chapter 3 Configuring, Plumbing, and Priming the Workstation

3-6 Applied Biosystems

Configuring

columns 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 Configuration

Dialog 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.

Column

position

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Thermo Fisher Scientific BioCAD® 700E Workstation Quick start guide

Brand: Thermo Fisher Scientific

Model: BioCAD® 700E Workstation

Category: Software manuals

Type: Quick start guide

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Thermo Fisher Scientific BioCAD® 700E Workstation Quick start guide

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