Dip and read™ Anti-GST Biosensors

Dip and Read Anti-GST Biosensors
Dip and Read™ Anti-GST Biosensors
For Quantitation and Kinetic Characterization of GST-tagged Biomolecules
Key Features
• Direct and rapid quantitation of GST-tagged biomolecules
• Easy capture of GST-fusion proteins for kinetic analysis
• Designed for use in buffer and complex media
• Compatible with Octet and BLItz platforms
Overview
Glutathione-S-transferase (GST) is commonly fused to recombinant proteins to facilitate detection and purification, and increase
solubility. The robust properties of the GST tag have established it
as the standard platform for protein-protein interaction pull-down
assays. The Anti-GST biosensor consists of a high affinity anti-GST
antibody pre-immobilized on a Dip and Read biosensor. It pro-
vides a rapid and label-free method for both quantitation of GST
targets (Figure 1) and kinetic analysis of GST fusions and interaction partners (Figure 2). The high specificity of the antibody-based
biosensor enables direct analysis of GST analytes in crude lysates,
column eluents, cell lysates and cell culture supernatants, serving
as a time-saving alternative to traditional analytical methods.
A
B
2.6
2000
1000
500
2.4
200
100
50
2.5
1
0.5
25
10
5
1.000
0.25
Standard
Fitting curve
2.2
2.0
1.8
0.100
Binding Rate
Binding, nm
1.6
1.4
1.2
1.0
0.010
0.8
0.6
0.4
0.2
0.0
0
20
40
60
T ime (sec)
80
100
120
0.001
1.0
10
100
1000
Concentration (µg/mL)
FIGURE 1: Quantitation measurement of a GST fusion protein on an Octet RED system showing concentration series of analyte (A) and resulting standard curve (B).
1
Dip and Read Anti-Mouse IgG Fc Capture Biosensors
EXIBILITY AnD VERSATILiTY
0.7
0.6
0.5
Binding, nm
The Anti-GST Dip and Read biosensor represents a new product
class qualified for both quantitation and kinetic applications.
Researchers will now be able to quickly quantify a GST fusion
protein and subsequently perform rigourous kinetic characterization of the fusion protein with an interaction partner.
Combined with the ease of use provided by Octet instruments,
these biosensors minimize and potentially eliminate sample
preparation. Compatibility with the BLItz platform further enables both quantitation and kinetic low volume applications.
For kinetic screening and characterization measurements,
Anti-GST biosensors can be regenerated up to 10 times using a standard low-pH protocol (see Tech Note 29, Anti-GST
Biosensor Kinetics Assays). Regeneration detaches the captured
ligand and bound analyte, clearing the surface for subsequent
analyses. For best results in rigorous kinetic applications, using
a fresh Anti-GST biosensor for each unique capture ligand is
recommended.
0.4
0.3
0.25 nM
0.031 nM
0.125 nM
0.0156 nM
0.2
0.1
0.0
0
300
600
900
1,200
1,500
1,800
2,100
2,400
T ime (sec)
FIGURE 2: Kinetic analysis of the interaction between GST-Protein A (65 kDa)
and an interaction partner Protein B (40 kDa) using an Octet RED system. 10X
kinetics buffer was used as the matrix throughout and the assay temperature
was 30°C.
RANGE OF APPLICATIONS
Anti-GST biosensors offer unprecedented ease of use and time
to- result for a wide variety of applications that include:
• Rapid quantitation of GST fusions (purified and crude)
• Kinetic characterization of interactions between GST fusions
and binding partners (purified and crude)
• Complete kinetic profiles and off-rate screening
Cycle
kon
koff
kD
1
1.5x104
1.9x10-5
1.2 nM
2
8.5x103
4.0x10-5
4.7 nM
3
6.8x103
1.810-5
2.6 nM
Table 1: Kinetic results for the interaction between GST-Protein A and Protein
B over three cycles using Anti-GST biosensors. Anti-GST biosensors were
regenerated using glycine pH 1.7 between cycles.
• Easy epitope binning/mapping
• Cell line monitoring in bioprocessing
• Upstream fermentation
• Downstream harvest and purification
• Process development
• Quality control
ORDERING INFORMATION
Order No.
733-2144
Description
Anti-GST (GST)
Biosensor for BLItz™
Qty.
1 Tray à 96 pcs.
CONTACT INFORMATION
PEQLAB Biotechnologie GmbH
Carl-Thiersch-Str. 2 B
91052 Erlangen, Germany
+49 9131 617 70 - 20
Freecall (D): 0800 100 20 16
[email protected]
www.peqlab.com
ForteBio,Inc.
ForteBio,
Inc.
1360 Willow Road, Suite 201
Menlo park,
Park, cA
CA 94025
t: 888.Octet-Qk
888.OCTET-75
or 650.322.1360
ForteBio, UK, Ltd.
ForteBio,UK,Ltd.
83 Victoria Street, Suite 407
London, SW1h
SW1H 0HW,
0hW, UK
Uk
t: +44-(0)20-31784425
www.fortebio.com
ForteBio
ForteBio(AriaBiotechnologyCo.Ltd.)
(Shanghai) Co., Ltd.
917 halley
Halley Road, Bldg 4
Zhangjiang high
High tech
Tech park
Park
Shanghai, china
China 201203
t: +86-21-51320387
© 2011 ForteBio, Inc. ForteBio, Octet, Dip and Read,
2010
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Inc. ForteBio
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and Read,and/or
and the
aredip
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