Embed Size (px)
Citation preview
BIOLOGICS SAMPLE PREPARATIONINTELLIGENTLY AUTOMATED
Agilent AssayMAP Platform
The Agilent AssayMAP Bravo platform.
SIMPLIFY BIOLOGICS SAMPLE PREPARATION
The Agilent AssayMAP platform is an open access, walkaway automation solution specifi cally designed for biomolecule sample preparation. It is based on the powerful combination of miniaturized, packed-bed chromatography cartridges, the state-of-the-art Bravo Automated Liquid Handling Platform, and a simple, application-based user interface that creates an open access environment for both novice and experienced automation users.
With a broad range of chemistries and laboratory-tested protocols that can be fl exibly combined, the AssayMAP platform simplifi es the most challenging sample preparation workfl ows.
• Perform quantitation and sample preparation in the same workfl ow • Reduce errors and increase walkaway time with reliable, automated processes • Increase throughput using existing chemistry or customized workfl ows
The AssayMAP platform is based on three key elements:
• Microchromatography cartridges with a wide range of standard chemistries
• The Bravo Automated Liquid Handler equipped with proprietary, positive-displacement, probe syringes
• User-friendly VWorks Automation Control software designed for simplicity in an open access environment
Versatile Resin CartridgesAssayMAP cartridges incorporate a 5 µL packed bed of resin supported by membranes molded into the polypropylene cartridge, enabling bidirectional fl ow. The cartridges accommodate virtually any polymer, silica, or gel-based resin with particle diameters from 15 to 100 µm. Ready-made cartridge options include protein A (PA-W), protein G (PG-W), and streptavidin (SA-W) for purifi cation and quantitation, and trypsin (TR-W) and reversed-phase (RP-W) versions for peptide mapping applications.
Precise Flow Control SyringesLiquid fl ow is precisely controlled by zero dead volume, positive displacement probe syringes which connect with the cartridges. Flow rates may be set low enough to achieve quantitative binding or elution in a single pass, in either aspirate or dispense mode. Recoveries approach 100 % for most applications, with reproducibility similar to HPLC (< 5 % CV). The probe syringes are mounted in the special 96AM head used on the Bravo Automated Liquid Handler. They can also mount pipette tips for standard liquid handling operations.
Intuitive Flexible InterfaceThe AssayMAP platform includes a growing library of application-specifi c protocols that use simple, fl exible forms-based control. Assay developers can easily set key parameters without the need for an on-site automation engineer. Once a method is optimized, the forms can be customized for error-free, push-button routine operation or combined for more complex workfl ows. AssayMAP protocols are built using the proven Agilent VWorks software platform, enabling seamless integration with larger automation systems.
The AssayMAP platform provides the tools needed to purify biomolecules from cell lysates, cell culture supernatants, and serum, enabling analysis by plate readers, HPLC, CE, and LC/MS. Workfl ow operations include affi nity purifi cation, enzymatic modifi cation, chemical reactions, fractionation, concentration, and cleanup.
POWERFUL MODULAR TOOLS FOR COMPLEX WORKFLOWS
Immobilize Bind
N-Glycan Analysis Sample Preparation An AssayMAP method for N-glycan sample preparation has been developed in collaboration with ProZyme, Inc. This new automated method reduces the time required for the workfl ow from 3 days to 4 hours. The target glycoprotein is purifi ed, denatured, immobilized on a cartridge, and then digested with PNGase F that specifi cally releases N-glycans from the target protein. Next, the released glycans are chemically labeled with a fl uorescent dye. Excess dye and reactants are removed with a cleanup cartridge prior to analysis by HPLC, CE, or LC/MS.
Immobilize Digest
Combined Purifi cation and QuantitationAssayMAP cartridges quantitatively purify target proteins from complex samples such as cell culture supernatants, cell lysates, and serum or plasma samples for the development, manufacturing, and clinical testing of new biotherapeutics. Purifi cation of the target protein is quantitative so the purifi ed protein can be used to both calculate its concentration in the complex sample and for downstream processing in preparation for a wide range of analytical methods. Only AssayMAP technology enables the combination of individual, often laborious, operations into one continuous, high-precision, high-throughput workfl ow. Resins conjugated to streptavidin, protein A, and protein G are available for easy adaptation of your existing purifi cation workfl ows.
Bind Wash
SA-W
Purify Denature, Reduce, and Alkylate
Peptide MappingA wide range of applications involve the LC/MS analysis of proteins, including characterization of post-translational modifi cations of biotherapeutics, quantitation of low-abundance protein biomarkers, and proteomics research. These applications require a complex series of sample preparation steps, including affi nity purifi cation of the analytical target, digestion with enzymes such as trypsin, concentration, desalting, cleanup, and pre-fractionation of the peptide mixture. AssayMAP modules have been developed for each of these operations, so you can create combinations that meet the specifi c application requirements of your end-to-end automated workfl ow.
STREAMLINED SAMPLE PREPARATION FOR A GROWING RANGE
Glycan release and labeling using the N-glycan sample preparation workfl ow.
Label
10
10
0.0
30Minutes
RFU
AnalyzePurify
Purifi cation of an antibody using protein A or protein G.
Affi nity purifi cation of an antigen using a streptavidin-biotin immuno-capture workfl ow.
pH 2
Elute
Elute
1.2
0.00 3000Concentration
(µg/mL)
A 280
AnalyzeWash
1.2
0.00 3000Concentration
(µg/mL)
A 280
Analyze
Purifi cation, proteolysis, and desalting of a target protein using the peptide mapping workfl ow.
TR-W
Enzyme Digest
0 30Minutes
100
0
A 215
Analyze
RP-S
Peptide Cleanup
OF APPLICATIONS
dashed rule = fold
Learn morewww.agilent.com/lifesciences/assaymap
Reach a local automation product specialist1-866-428-9811
USA and [email protected]
Asia Pacifi [email protected]
For Research Use Only. Not for use in diagnostic procedures. This information is subject to change without notice.
© Agilent Technologies, Inc., 2012Printed in the USA, July 12, 20125991-0668EN
