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Agilent’s NEW AssayMAP Bravo
A High Throughput Microchromatography Platform
for Quantitative Analytical Scale Protein Sample Preparation
The Agilent Bravo® Liquid Handler
Best in Class Compact Liquid Handling• High accuracy pipetting from
• 300nL to 250μL into 96, 384 and 1536
• Whole Plate, Column/Row-wise, Single Tip Transfer pipetting
• Can use for example Eppendorf Cups, 24 well cell culture plates etc ....
• head change in 30s (96 to 384 or AssayMAP head for example)
• On deck plate stacking & tip nesting (with gripper)
• Re-use of tips up to 100 times (Washing station available)
Adapt the Deck to Nearly Any Application• On-deck barcode reading, plate heating & cooling, shaking.
• Auto-filling reservoir, vacuum locating of warped PCR plates.
• Magnetic bead separations, vacuum filtration.
Applications
All liquid handling tasks like: serial dilution, normalisation, hitpicking, cell culture, ELISA, screening, genomics (DNA/RNA-Extraction, NGS, PCR ...), InGelDigest, ...
..... many others more ...
The AssayMAP Bravo
The AssayMAP Bravo is a solution forquantitative, analytical scale proteinsample preparation to deliver highlyparallel microscale purification tobiologics discovery & development.
The AssayMAP Bravo enables workflows in biologics discovery & development for…
• MAb titer and characterization including cell line selection & cell culture optimization
• rapid ELISA
• protein capture and purification
• chromatography method development.
Page 3
One pot. Bottleneck in Protein Sample Processing
Manufacturingprocess
Biologicalsystem
Clinicalpatient
Samples
Mass spectrometer
HPLC
Plate reader
Instruments
AssayMAP Bravo Head & Syringe
Bravo 96AM head with AssayMAP technology
• Direct displacement, ultra-low dead volume syringe with probe.
• Liquid handling via probes directly, through the cartridge.
• Unidirectional liquid handling through cartridges for fine control of separation and true, single pass, chromatography.
• Fully enabled for Bravo liquid handling – pipette tips can be pressed on the probe, partial head, deck accessories, ….
AssayMAP Cartridges
Chromatography column on a micro scale.
• Various chromatographic resin as packed beds between two supports
• Can be affinity resins or immobilized enzymes
• IEX, RP, HIC, HILIC, ….
• Streptavidin, protein A/G, lectin, metal chelates, ….
• Trypsin, PNGase, ….
AssayMAP vs. Pipett Tip Columns
AssayMAP Cartridge Pipett Tip Column
• Single entry/exit port
• Multi-pass contact (equilibrium adsorption)
• Less controllable flow rate
• Air entrainment
• Non-quantitative extraction
• Separate entry & exit ports
• Single-pass contact (“true chromatography”)
• Controlled flow rate
• No air entrainment
• Fully quantitative binding & elution
AssayMAP Bravo VWorks Software
Page 9
VWorks
Flow rate control.
VWorks Task Macros for AssayMAP Tasks
AssayMAP Bravo - VWorks Data entry form
Page 10
VWorks Form
VWorks Software features for AssayMAP protocols.
• VWorks forms & protocols packaged with each AssayMAP application.
• User specified flow rate control to fine tune AssayMAP parameters. Fast flow to equilibrate, slow flow to bind sample.
AssayMAP Bravo MAb Purification on Protein A
Quantitative recovery, excellent purity & reproducibility with no detectable carryover.
AssayMAP Toolkit
40 µm glass beads
Non-porous
~1 µg IgG coating capacity
Streptavidin (SA)
CaptureSelect®
Ligands (human Fc)
Protein A (PA)
HydrophobicBinding Surface (RP-C18)
Wide Pore (W)
Surface Chemistries Bead Types
Non-Porous (N)
40 µm polymer beads
Perfusive wide pore
~100 µg IgG coating capacity
Available Now
Still to come: IEX, other hydrophobic, IMAC/metal chelaters, other affinity (lectins, protein G, …), immobilized enzyme (proteases, glycosidases…) and custom resins.
Example: Rapid ELISA
Micron scale diffusion path
15 – 30 minutesturnaround
Millimeter scale diffusion path
4 – 24 hoursturnaround
Microplate
AssayMAP
Coat Block Sample Conjugate Substrate Read
Protein Biomarker Quantitation for LC/MS
15
© Plasma Proteome Institute
Digestion Purification AnalysisPrefilter
Complete SISCAPA sample preparation on the AssayMAP Bravo.
In Dev In Dev Now!
AssayMAP - Bravo
• Affinity purification or bind/elute assays (e.g. cell culture titer, PK studies)
• ELISA (e.g. process impurity or biomarker assays)
• Sample preparation for characterization.
• Chromatographic method development
• 5 µL packed bed cartridge in 96-channel ANSI/SBS microplate format
• Capacity per preparation up to ~100ug
• Single pass, fully quantitative binding & elution
• Positive displacement, bidirectional flow control
• Quantitative recovery, excellent purity & reproducibility with no detectable carryover
High throughput microchromatography platform
Potential applications