Large Capacity Cartridge and Workbench 3.2 Overview
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.1
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.2
New Cartridge Size to Address More Workflows
Enable superior Mass Spec Results
with AssayMAP Bravo
5ul 25ul
May 21, 2020Title Confidentiality label Regulatory statement (if applicable)
Biopharma
Large Capacity Affinity Cartridges Enables More Parallel Analyses
Detect Analyze
Automated Sample Preparation
Peptide Mapping
Protein Quant
Functional Assays
Protein Aggregation
Intact & Fragment
PTM
Protein A
Streptavidin
Detect Analyze
May 21, 2020 Title Confidentiality label Regulatory statement (if applicable)4
Proteomics
Large Capacity Cleanup Cartridges Enables More Workflows
Detect
Phosphopeptide
Analysis
Analyze
Automated Sample Preparation
Untargeted Proteomics
Targeted Proteomics
GAP
C18
RPS
PAW Performance-Binding Capacity
PAW
Cartridge
Type
Quantitative
Binding
Capacity
5 µL 100 µg IgG
25 µL 800 µg IgG
25 µL
5 µL
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.5
PAW Performance-Reproducibility and Recovery
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.6
0
100
200
300
400
500
600
700
800
900
1000
100 300 500 700 900
hIg
G(µ
g)
elu
ted
hIgG (µg) loaded
Elution and Flow through
Flow-through Eluate
Sample Load (ug) %Recovery %CV
1-3 100 87 3.1
4-6 300 92 1.4
7-9 500 99 3.1
10-12 700 96 2.6
13-15 900 95 2.7
PAW Performance-Minimum Elution Volume
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.7
0
50
100
150
200
250
300
350
400
10 20 30 40 50 60 70 80 90 100 110
hIg
G(µ
g) e
lute
d
Cumulative elution volume (µL)
Elution Profile
500 ug of hIgG was loaded on large capacity PAW cartridges and eluted sequentially with 10 µL of elution solution.
93% recovery in 30 µL
SAW Performance-Binding Capacity
SAW
Cartridge
Type
Quantitative
Binding
Capacity
5 µL 50 µg B-BSA
25 µL 300 µg B-BSA
25 µL
5 µL
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.8
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.9
Application Updates to Support Large Capacity Cartridges
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.10
Software App/Utility Versioning
May 21, 2020 Agilent Confidential11
App X.YMajor Version Minor Version
X = Protocol change
Y = Form change only
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.12
App Library
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.13
Obsoleting
May 21, 2020 Agilent Confidential14
Grayed out applications are applications from Workbench 3.1 that have a
newer version in Workbench 3.2. Grayed out applications are still functional
but will be removed in the next workbench release.
Applications with a Name Change
Workbench 3.1 Workbench 3.2 Reason for Update
Antibody Purification v1.0 Affinity Purification: Aspiration Mode v2.0 Eluate Mixing added
Peptide Cleanup v1.1 Peptide Cleanup: Aspiration Mode v2.0 Eluate Mixing added
Aspiration Mode vs Non-Aspiration Mode
We offer both standard and aspiration mode protocols for Affinity Purification and Peptide Cleanup. However, we recommend the standard protocols for most users
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.17
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.18
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.19
New Application versions created to support large capacity volume
Workbench Changes
Method saving added to: Affinity Purification v2.0, Immobilization v2.0, Peptide Cleanup v3.0, Protein Cleanup v2.0
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.20
New Application versions created to support large capacity volume
Workbench Changes
Access default parameter settings for 5µL and 25µL cartridges via the Browse Method button
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.21
5µL Settings
5µL Cartridge Settings vs 25µL Cartridge settings
25µL Settings
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.22
Navigation Updates
Workbench Changes
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.23
Utility Library
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.24
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.25
Normalization Moved from App Library to Utility Library
Utility Changes
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.26
Updated syringe wash and drying steps at start of protocol
Major Revision Utilities
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.27
Navigation Updates
Utility Changes
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.28
Workflow Library
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.29
SISCAPA Workflow has been removed
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.30
Literature Library
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.31
Highlighted Guides are new. All App & Utility Guides have been updated
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.32
These Major Changes were Made
Major Changes
App/Utility/Workflow Change
Affinity Purification Method Saving/Large Capacity Support/Added Air Gap Prevention Step/Sample Chase changed from 5µL to 25µL
Immobilization Method Saving/Large Capacity Support/Added Air Gap Prevention Step/Sample Chase changed from 5µL to 25µL
Peptide Cleanup Method Saving/Large Capacity Support/Added Air Gap Prevention Step/Sample Chase changed from 5µL to 25µL
Protein Cleanup Method Saving/Large Capacity Support/Added Air Gap Prevention Step/Sample Chase changed from 5µL to 25µL
Affinity Purification: Aspiration Mode Re-Activated/Added Eluate Mixing
Peptide Cleanup: Aspiration Mode Re-Activated/Added Eluate Mixing
Normalization Updated Syringe Wash and Drying Steps
Reformatting Updated Syringe Wash and Drying Steps
Serial Dilution Updated Syringe Wash and Drying Steps
SISCAPA Workflow Removed
Peptide Sample Prep Workflow v1.0 Removed
Reagent Transfer v1.0 Removed
Sample Plate Transfer v1.0 Removed
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.33
These Minor Changes were Made
Minor Changes
App/Utility/Workflow Change
Fractionation
Updated Form image, Standardized navigation,
Standardized nomenclature
IMAC Cartridge Customization
In-Solution Digestion: Multi-Plate
In-Solution Digestion: Single-Plate
On-Cartridge Reaction
Phosphopeptide Enrichment
Cartridge Transfer
Pipette Tip Transfer
Reagent Aliquot
Reagent Transfer
Single Liquid Addition
Syringe Test
Syringe Wash
System Startup/Shutdown
Creating a Method
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.34
Open Default
Method
Modify parameters
to your needsName Method Save Method
Additional error handling has been implemented. See user guides for additional information
Note: This procedure is for: Affinity Purification v2.0, Immobilization v2.0, Peptide Cleanup v3.0, Protein Cleanup v2.0
Method Folder Location
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.35
Note: Methods files are not backwards compatible
Required Hardware Changes to use Large Capacity Cartridges
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.36
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.37
Wider Cartridge diameter requires new Chimneys to prevent crashes
Changes Wash Station
Note: For additional information, see the 96 Channel Wash Station Maintenance Guide at Agilent.com
Existing Customers
• Customers can do it themselves with upgrade instructions
• Instructions to Replace the chimneys in Wash Station Maintenance Guide
• Can be done through FSE, regular PM, or PS/AE
May 21, 2020 For Research Use Only. Not for use in diagnostic procedures.38
How do you get a Wash Station Upgraded
Wash Station Upgrade
Remove Chimneys
with needle-nose pliers
1 2
May 21, 2020 Title Confidentiality label Regulatory statement (if applicable)39
AssayMAP Community Page
Customer Resources