Bio-Rad ChromLab Software User Manual

3.0

NGC™ Chromatography Systems

and ChromLab™ Software

User Guide

Version 3.0

NGC™ Chromatography
Systems and
Chr

User Guide

Version 3.0

omLab™ Software

Bio-Rad Technical Support Department

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Table of Contents

Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11

Main NGC Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

NGC Chromatography Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Finding Out More . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

Chapter 2 The Workspace . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17

The Home Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

The System Control Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

The Method Editor Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

The Evaluation Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Instrument Control Touch Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Touch Screen Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Touch Screen Toolbar Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Chapter 3 System Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .29

System Control Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

View Menu Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Manual Menu Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Tools Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Toolbar Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Context Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

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Table of Contents

Chromatogram View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

Changing Chromatogram View Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Showing or Hiding Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39

Changing Trace Color . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Zooming In and Out. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Changing the X-Axis Units and Scale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Changing the Y-Scale Values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Autoscaling the UV Trace . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Annotating the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

Showing and Hiding Events, Fractions, Errors, and Annotations . . . . . . . . . 44

Fluidic Scheme Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

Modules. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Fluidic Scheme Configurations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

Working with Fluidic Schemes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .57

Fluidic Scheme Mapping. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Calibrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64

Calibrating a Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .64

Calibrating Flow Rate of Pumps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

Calibrating Pressure Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

Verifying Plumbing with the Point-to-Plumb Feature . . . . . . . . . . . . . . . . . . . . . . 67

System Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .69

Delay Volume Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69

Control Flow Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Remote Access Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .74

Trace Settings Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .74

Device Input Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76

Device Output Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

Air Sensors Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80

Email Notifications Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82

System Name Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85

System Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .86

General Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Devices Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Process Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

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Detector Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87

Single-Wavelength Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87

Multi-Wavelength Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88

Preferences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .89

Email Server Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89

Chapter 4 Performing a Manual Run. . . . . . . . . . . . . . . . . . . . . . . . . . 93

Preparing the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94

Priming and Purging the System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94

Cleaning the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .102

Running an Experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

Changing Module Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Stopping a Manual Run. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Clearing Run Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Saving a Manual Run. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Saving a Manual Run on the Touch Screen . . . . . . . . . . . . . . . . . . . . . . . . . . 110

Viewing Run Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111

Chapter 5 Method Editor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113

Before You Begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .114

Accessing Method Editor Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114

Opening a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115

Method Editor Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117

Edit Menu Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118

View Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .119

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119

Toolbar Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119

Phase Library Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .120

Adding a Phase to a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121

Method Outline Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122

Phase Parameters Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122

Method Steps Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123

Viewing Details of a Step in the Step Table . . . . . . . . . . . . . . . . . . . . . . . . . . 124

Adding, Modifying, and Deleting Steps in the Step Table. . . . . . . . . . . . . . . . 124

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Method Settings Parameters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125

Fluidic Scheme . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125

Column Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126

Detector Settings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131

Unit Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131

pH Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

Fraction Collection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

Flow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133

Buffer Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133

Phase Controls and Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136

Standard Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138

Sample Application Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138

Column Wash Parameters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143

Elution Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144

System CIP (Clean in Place) — Storage Parameters . . . . . . . . . . . . . . . . .145

Column Preparation/Activation Parameters . . . . . . . . . . . . . . . . . . . . . . . . 152

Column Performance Test Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . .153

System Preparation Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154

Create New Phase Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155

Step Library Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156

Scout Parameters Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157

Chapter 6 Creating a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159

Standard Method Templates. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160

Creating a Method from a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .163

Creating a Method from Standard Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165

Adding Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170

Editing Methods and Phases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170

Renaming Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171

Rearranging Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171

Deleting Phases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172

Opening a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172

Running a Method and Collecting Fractions. . . . . . . . . . . . . . . . . . . . . . . . . . . . 173

Collecting Fractions Using the BioFrac Fraction Collector . . . . . . . . . . . . .173

Collecting Fractions Using Outlet Valves . . . . . . . . . . . . . . . . . . . . . . . . . .173

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Collecting Fractions in Run Queues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174

Collecting Fractions Using the BioFrac Fraction Collector . . . . . . . . . . . . .174

Collecting Fractions Using Outlet Valves . . . . . . . . . . . . . . . . . . . . . . . . . .174

Saving a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .177

Renaming a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178

Deleting a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179

Scouting on Parameters within a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180

The Scouting Wizard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .181

Page 1: Choose a Scouting Parameter . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181

Page 2: Select Method Steps to Scout . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182

Page 3: Generate Scout Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184

Using the Scouting Wizard to Scout Columns . . . . . . . . . . . . . . . . . . . . . . 185

Using the Scouting Wizard to Scout Samples . . . . . . . . . . . . . . . . . . . . . . 187

Creating a Scouting Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189

Special Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190

Viewing a Summary of the Scouting Method . . . . . . . . . . . . . . . . . . . . . . . . . 192

Running a Scouting Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192

Chapter 7 Evaluating Results. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195

Managing Analysis Projects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196

Evaluation Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 198

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199

View Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .201

Analysis Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 202

Tab Toolbar Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 202

Customizing the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203

Displaying Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203

Selecting the Active Trace. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204

Showing or Hiding a Trace . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205

Changing the Axes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .205

Controlling the Range of the Y-Scale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205

Changing Trace Colors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206

Zooming In and Out. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207

Obtaining Statistics of a Selected Region on the Chromatogram . . . . . . . . . 208

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Annotating the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210

Copying the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211

Customizing the Trace Table Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 212

Changing Table Grouping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 212

Sorting Table Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213

Ordering and Selecting Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213

Showing or Hiding Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213

Copying a Table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214

Showing or Hiding Pre-Injection Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214

Manually Setting the Injection Point (x Alignment) . . . . . . . . . . . . . . . . . . . . . 215

Managing Runs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216

Renaming a Single Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216

Deleting a Single Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216

Viewing Multiple Runs in Separate Tabs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 217

Comparing Traces. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218

Viewing Multiple Runs in One Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219

Stacked View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219

Overlay View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221

Customizing the Trace Comparison Table . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Expanding or Collapsing Groups. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Hiding All Traces in a Group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Showing All Traces in a Group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Offsetting Traces — Overlay Mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223

Managing Analyses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .224

Removing Runs from a Trace Comparison . . . . . . . . . . . . . . . . . . . . . . . . . . . 224

Saving a Trace Comparison Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 224

Copying an Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225

Renaming an Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .225

Deleting an Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .226

Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226

Exporting Run Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226

Peak Integration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .227

Automatic Peak Integration Settings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

Baseline Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

viii | NGC Chromatography Systems and ChromLab Software

Table of Contents

Peak Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .229

Peak Filtering . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229

Default Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229

Integrate and Delete . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .230

Starting Peak Integration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230

Displaying Peak Results — the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . 231

Changing Peak View Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232

Displaying Peak Results — the Peaks Table. . . . . . . . . . . . . . . . . . . . . . . . . . 233

Table Display Order and Column Selection . . . . . . . . . . . . . . . . . . . . . . . . 235

Showing or Hiding Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235

Copying the Peaks Table. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236

Grouping or Ungrouping the Peaks Table. . . . . . . . . . . . . . . . . . . . . . . . . . 236

Clearing Extinction Coefficient Values in the Peaks Table . . . . . . . . . . . . . 237

Adjusting Peaks Manually . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 238

Saving Peak Integration Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 240

Evaluating Fractions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241

Displaying Fraction Collection Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241

Displaying Fraction Collection Results — the Fractions Table . . . . . . . . . . . . 243

Calculating Protein Concentration for Fractions . . . . . . . . . . . . . . . . . . . . .245

Viewing Fraction Details . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .246

Pooling Fractions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246

Viewing Fraction Collection Results — the Heat Map. . . . . . . . . . . . . . . . . . . 248

Viewing Heat Map Details . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 249

Column Performance Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251

Column Performance — Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251

Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 252

Calculate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 252

Analyzing Column Performance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 253

Resetting the Column Performance Properties. . . . . . . . . . . . . . . . . . . . . . 254

Deleting Column Performance Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254

Column Performance — Manual Adjustment . . . . . . . . . . . . . . . . . . . . . . . . . 255

Displaying Column Performance Results — the Chromatogram . . . . . . . . . . 255

Displaying Column Performance Results — the Column

Performance Table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256

User Guide | i x

Table of Contents

Chapter 8 Importing and Exporting Data . . . . . . . . . . . . . . . . . . . . . 259

Importing an NGC Method or Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 259

Importing Unicorn Data Files. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261

Importing BioLogic DuoFlow Data Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 263

Exporting Data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264

Exporting Data as an NGC File . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 265

Exporting Run Data as a .csv File . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 269

Exporting Diagnostic Logs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .271

Chapter 9 Reports. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275

Producing a Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276

Method Reports. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277

Run Reports. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277

Analysis Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278

Printing a Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278

Saving a Report. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 279

Appendix A Database Management . . . . . . . . . . . . . . . . . . . . . . . 281

Back Up the NGC Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .282

Restore the NGC Database. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 283

Appendix B Multicolumn Purifications . . . . . . . . . . . . . . . . . . . . . 285

Multicolumn Purification Method Templates. . . . . . . . . . . . . . . . . . . . . . . . . . . . 286

Multicolumn Sequential Purification Templates . . . . . . . . . . . . . . . . . . . . . . . 286

Multicolumn Tandem Purification Templates . . . . . . . . . . . . . . . . . . . . . . . . . 289

Performing Multicolumn Purification Chromatography. . . . . . . . . . . . . . . . . . . . 291

Plumbing the NGC System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291

Plumbing NGC Discover Pro Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291

Plumbing the NGC Discover Pro System to Use

Multicolumn Purification Templates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .292

Priming the NGC System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 294

Selecting and Saving a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295

Running Multicolumn Purification Methods . . . . . . . . . . . . . . . . . . . . . . . . . . 296

x | NGC Chromatography Systems and ChromLab Software

1 Introduction

NGC™ chromatography systems provide a general purpose purification platform for
purifying all forms of biomolecules using a combination of chromatography
techniques. The systems are also useful for developing and optimizing purification
protocols. The systems can provide highly purified proteins, peptides, nucleic acids,
monoclonal antibodies, and other small molecules.

User Guide | 1 1

1 | Introduction

ChromLab™ software enables you to set up and control an NGC instrument, run
protein separations and other operations manually, program methods to automate
purification runs, evaluate the results, and generate and print experiment reports.
This user guide explains how to perform all these tasks.

Main NGC Features

NGC chromatography systems enable you to do the following:

 Easily create purification and maintenance protocols from predefined

method templates and protocol phases

 Automate multicolumn purification processes using preprogrammed

templates and multiple column switching valves

 Automate multiple sample injections using either the sample inlet valve and

the sample pump or the C-96 autosampler

 Expand sample monitoring using the signal import module (SIM) to export

digital signals to and import digital signals from external detectors

 Collect large-volume fractions using multiple outlet valves while also

collecting small-volume fractions using the BioFrac™ fraction collector

 Automatically prepare buffers using preprogrammed buffer blending

protocols

 Analyze purification results through 1-click peak integration, determine

protein concentration and calculate column performance

 Automate purification protocol optimization using the scouting wizard

 Easily locate fractions containing peaks of interest and view the protein

concentration within each fraction

 Extend the preconfigured systems with additional valves for buffers,

samples, and columns

12 | NGC Chromatography Systems and ChromLab Software

Main NGC Features

 Organize the location of the modules to optimize separation performance

based on method scale and complexity, and to minimize the system swept
volume

 Minimize errors when connecting tubing using the Point-to-Plumb™

feature in ChromLab software

User Guide | 1 3

1 | Introduction

NGC Chromatography Systems

All NGC chromatography systems include ChromLab software and the NGC touch
screen.

NGC chromatography systems are available in several combinations. Each system
is equipped with either two 10 ml system pumps (the 10 series) or two 100 ml
system pumps (the 100 series).

The NGC Quest™ chromatography system includes the following:

 Two system pumps

 Mixer

 Sample injection valve

 Conductivity monitor with either a single-wavelength UV detector or a

multi-wavelength UV/Vis detector (available on the Plus systems)

14 | NGC Chromatography Systems and ChromLab Software

NGC Chromatography Systems

The NGC Scout™ chromatography system includes the following:

 All modules on the Quest system

 pH detector valve

 Buffer blending valve

The NGC Discover™ chromatography system includes the following:

 All modules on the Scout system

 Column switching valve

 Two buffer inlet valves

 Sample pump

The NGC Discover Pro chromatography system includes the following:

 All modules on the Discover system

 Fourth expansion tier

 Sample inlet valve

 Outlet valve

User Guide | 1 5

1 | Introduction

Finding Out More

After you install NGC documentation from the NGC Chromatography Systems
Documentation DVD, you can access installed NGC guides and tutorials on the
Help menu in any ChromLab view.

More information about the NGC chromatography systems and ChromLab software
is available fr

 The NGC Chromatography Systems and ChromLab Software Installation

Guide is available on your NGC Chromatography Systems Documentation
DVD as a .pdf file. This guide explains how to set up your environment, set
up and install the NGC instrument in the lab, install ChromLab software,
and connect ChromLab to the NGC system.

 The NGC Chromatography Systems and ChromLab Software Instrument

Guide is available on your NGC Chromatography Systems Documentation
DVD as a .pdf file. This illustrated guide details the modules that make up
the NGC instrument and includes troubleshooting and maintenance
information.

 For ChromLab Help, clicking in dialog boxes accesses relevant

information. Screen-level help is also available on the Help menu.

om the following sour

ces.

 NGC video tutorials are available on the NGC Chromatography Systems

Documentation DVD as .mp4 files.

Tip: You can click the Bio-Rad logo in the upper-left corner of any
Chr

omLab window to launch the Bio-Rad website.

16 | NGC Chromatography Systems and ChromLab Software

2 The Workspace

ChromLab™ software provides an intuitive interface for developing chromatography
methods, operating an NGC™ instrument, and analyzing data from chromatography
runs.

ChromLab software presents four primary workspaces.

 The Home window

 The System Control window

 The Method Editor window

 The Evaluation window

Each workspace is shown and described in this chapter. The NGC instrument touch
scr

een is also described.

User Guide | 1 7

2 | The Workspace

The Home Window

ChromLab software opens with the Home window, which displays three panes and
the System Control tab.

The three panes provide quick access to the system control, method editor, and
evaluation workspaces. The status of connection to the NGC instrument is also
displayed. Links to recently completed runs, recently created methods, and recently
accessed or analyzed runs and analyses appear listed at the bottom of the window.

Tip: Clicking the Bio-Rad logo in the upper-right corner of any ChromLab
window launches the Bio-Rad website.

18 | NGC Chromatography Systems and ChromLab Software

The Home Window

Method Editor and Evaluation tabs become visible in the Home window when you
select tasks in the Method Editor and Evaluation panes.

User Guide | 1 9

2 | The Workspace

File Menu Commands

Connect to System — opens a dialog box that enables you to choose an NGC

chromatography system to connect to. ChromLab detects the NGC systems
available on the same subnetwork or those systems that are directly connected to
the computer. Their system name, network name, and IP address appear in the
table. To connect to a system, you can:

 Select a name in the list of detected systems and click Connect.

 Select the appropriate radio button, enter the system’s name or IP

Note: If your system does not appear in the list, click Detect.
Chr

omLab searches the network for available NGC systems and

refreshes the list.

address, and click Connect.

Tip: To obtain the system’s name and IP address select System
Information on the instrument touch scr

een dropdown menu.

Disconnect System — starting Chr

omLab connects you to the NGC system. This
command enables you to disconnect ChromLab software from the system so you
can work offline.

Manual Run — opens the System Contr

ol window in manual mode so you can

perform a manual run or set up your system manually.

Calibrate — opens the Calibration dialog box, which displays instructions and
options for selecting a module and calibrating it. See Calibrations on page 64 for
details.

Open Template — opens the T

emplate dialog box in which you can select a

method template from template folders organized by technique.

Open Method — opens the Method dialog box in which you can select fr
your saved methods.

New Method — opens the Method Editor window in which you can cr
method using standard method phases and steps.

20 | NGC Chromatography Systems and ChromLab Software

om a list of

eate a

The Home Window

Import — displays links from which you can import the following:

 NGC File — opens a dialog box in which you can import a method, a

method with runs, or a run exported from ChromLab software running on
another NGC system. See Importing an NGC Method or Run on page 259
for more information.

 Unicorn Data — opens a dialog box in which you can import a Unicorn

data file into the NGC database. See Importing Unicorn Data Files on

page 261 for more information.

 DuoFlow Data — opens a dialog box in which you can import a BioLogic

DuoFlow™ data file into the NGC database. See Importing BioLogic

DuoFlow Data Files on page 263 for more information.

Export — displays links fr

 Methods/Method Runs — opens a dialog box in which you can export

om which you can export the following:

both single or multiple methods and single or multiple methods with
associated runs.

 Runs — opens a dialog box in which you can export single or multiple

runs.

See Exporting Data as an NGC File on page 265 for more information.

Open Run/Analysis — opens a dialog box in which you can select a run or analysis
to view or analyze.

Preferences — opens dialog boxes in which you can do the following:

 Select pressure units for all system and software pressure values. This is a

global setting.

 Set up an SMTP server to receive email messages about system

notifications from the ChromLab computer.

Exit — closes Chr

omLab.

User Guide | 2 1

2 | The Workspace

Help Menu Commands

Help — displays screen-level help topics and links to installed manuals.

Export Diagnostic Logs — opens the Export Diagnostic Logs dialog box in which

you can export all critical information that Bio-Rad T
diagnose issues. The log files and data are zipped and saved to a location that you
choose. See Exporting Diagnostic Logs on page 271 for more information.

echnical Support requires to

About — displays Chr

omLab copyright and version information.

22 | NGC Chromatography Systems and ChromLab Software

The System Control Window

The System Control window enables you to run the instrument manually, monitor
method runs while they are running, select fluidic schemes, calibrate pumps and
detectors, and verify the accuracy of instrument plumbing. This window displays a
chromatogram during a run. A fluidic scheme graphically depicts the flow path of all
the modules on the system. A status panel appears above each module displaying
its real-time status. In manual mode, clicking a module displays its controls and
detailed settings. The Run Log documents each action that occurs during a run. The
Run Queue lists all runs ready to be started.

The System Control Window

System Control functionality is detailed in Chapter 3, System Control.

User Guide | 2 3

2 | The Workspace

The Method Editor Window

The Method Editor window enables you to create, open, review, edit, and run a
method. You can also open and edit a method template to create a new template.
Method Editor functionality is detailed in Chapter 5, Method Editor. See also

Chapter 6, Creating a Method.

24 | NGC Chromatography Systems and ChromLab Software

The Evaluation Window

The Evaluation window enables you to view and compare run data, perform peak
integration, and save run data as analyses. Evaluation functionality is detailed in

Chapter 7, Evaluating Results.

The Evaluation Window

User Guide | 2 5

2 | The Workspace

Instrument Control Touch Screen

In addition to ChromLab software running on a computer, the instrument is
equipped with a touch screen that accesses system control functionality. You can
use this touch screen to run, control, and monitor a run independent of ChromLab.
See System Control on page 29 for more information.

Tip: When the NGC system has been inactive for two hours the LED display
scr

eens on the instrument tur
appears on the touch screen informing you that the system is in standby mode.
You can take the system out of standby mode by touching OK in the dialog
box, starting the system pumps by initiating a manual or method run, or clicking
on a module in the fluidic scheme that has an LED display.

n off, the touch screen dims, and a dialog box

26 | NGC Chromatography Systems and ChromLab Software

Instrument Control Touch Screen

Touch Screen Menu Commands

Calibrate — opens the Calibration dialog box, which displays instructions and

settings for selecting a module and calibrating it. See Calibrating a Module on

page 64 for details.

Point-to-Plumb — starts the Point-to-Plumb™ featur
instrument LED lights so you can visually verify or change instrument plumbing.
Displays the current fluidic scheme. See Verifying Plumbing with the Point-to-Plumb

Feature on page 67 for details.

Change Fluidic Scheme — opens the Fluidic Scheme Selector dialog box in which
you can edit the fluidic scheme or choose another one. See Fluidic Scheme

Configurations on page 51 for details.

Map Fluidic Scheme — opens the Fluidic Scheme Mapping dialog box in which
you can map devices on your instrument to their position in the fluidic scheme. See

Fluidic Scheme Mapping on page 60 for details.

System Settings — opens the System Settings dialog box in which you can
customize system settings. See System Settings on page 69 for more information
about customizing your system.

System Information — opens the System Information dialog box, which lists the
serial number and other general information about the NGC device as well as
information about the system components, pr
detectors.

Service — for Bio-Rad technical service staf

Help — displays detailed information about touch scr

ocesses, and UV and UV/Vis

f use only. Do not select this command.

e and simultaneously turns off

een menu commands.

About — displays Chr

Shut Down — shuts down the NGC system, including the connected computer

omLab version and copyright information.

User Guide | 2 7

.

2 | The Workspace

Touch Screen Toolbar Commands

Save — saves in a data file the steps executed during a manual run.

Clear — deletes manual run data fr

Autoscale — automatically scales the chr

peak height during the run. Autoscaling is enabled by default.

Reset Zoom — r

Annotate — adds a note to the chr

or after the run completes.

esets the view to show the full chromatogram.

om the touch screen display.

omatogram’s primary y-axis to the tallest

omatogram at points on the x-axis during a run

28 | NGC Chromatography Systems and ChromLab Software