A High Throughput Microchromatography Platform - Agilent · A High Throughput Microchromatography Platform ... Single Tip Transfer pipetting ... High throughput microchromatography

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.

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What is the AssayMAP Microchromatography Technology?

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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

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VWorks

Flow rate control.

VWorks Task Macros for AssayMAP Tasks

AssayMAP Bravo - VWorks Data entry form

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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

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

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© Plasma Proteome Institute

Digestion Purification AnalysisPrefilter

Complete SISCAPA sample preparation on the AssayMAP Bravo.

In Dev In Dev Now!

AssayMAP Bravo MAb Purification on Protein A -literature

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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

Question and answers?

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This information is subject to

change without notice

© Agilent Technologies, Inc. 2011

Published in Germany, July 2011