15 Minute universal Multiplex assay platforM

15 Minute
Universal
Multiplex Assay
Platform
BREAKTHROUGH UNIVERSAL MULTIPLEXING
FOR FAST, EASY & ACCURATE TESTING
The Maverick Detection System performs multiplexed label-free detection of protein binding events by
measuring wavelength resonance shifts caused by surface binding on silicon photonic sensors,
eliminating the need for fluorescent, luminescent, or radioactive labels used in other immunoassay formats.
The Maverick Detection System is designed around a proprietary disposable consumable – the
AutoArray – which contains 12 silicon chips, each with an array of patented Microring Sensors on its surface.
The binding of proteins or antibodies to any of the microring sensors on the chip is detected in
real-time and the results are simultaneously analyzed with accompanying Genalyte software.
4 Independent
Sensors per Analyte
nne
Cha
ple
Sam
Sa
mp
le
Ch
l1
an
ne
l2
16 Analytes per
Sample Channel
2 Sample Channels
per Chip
Each of the 12 chips in the AutoArray contain
two independent sample channels.
Each sample channel contains 16 sensor
spots, enabling up to 24 samples to be
tested against 16 analytes, or 12 samples
tested against 32 analytes per kit.
THE MAVERICKTM ADVANTAGE
SPEND TIME DOING RESEARCH NOT ASSAYS
Simple & Easy To Use
SMALL SAMPLE VOLUME
Automated workflow requires minimal
hands-on time and reduces or eliminates the
multiple washes, incubations, or additional
steps typical of other technologies.
On-board fluidics ensure assays are performed the
same way every time, reducing the opportunity for
variation and thereby lowering CV%.
5-10µL sample volume extends
the use of precious samples.
RESULTS IN ONLY 10 MINUTES
First results are reported in as little as 10 minutes.
That’s over 10x faster than traditional ELISA and
light speed ahead of bead-based technology.
LABEL-FREE TECHNOLOGY
Enables kinetic screening and affinity
measurements of multiple interactions.
THE MAVERICKTM SILICON PHOTONIC CHIP
The key to the Maverick Detection System is its patented silicon chip containing arrays of
Microring Sensors that can simultaneously analyze multiple proteins from a single sample.
HOW IT WORKS
1. Each microring sensor on the chip is
coated with a distinct antigen or
antibody which enables multiplexing.
2. An infrared laser generates light in a
broad wavelength range which traverses
waveguides on the chip surface.
3. Each ring resonates at a particular wavelength
causing only that wavelength to be trapped in
the ring while all others continue on to a
photodetector. This results in a measurable “notch”
in the spectrum detected by the instrument.
4. As material binds to the sensor surface the wavelength of required for resonance
within the ring changes, causing the “notch” to move (resonance shift).
5. The resonance shift changes proportional to the amount of material bound to the ring surface.
This shift is monitored in real-time by the Maverick instrument and results plotted in picometers
of wavelength shift over time - Genalyte Response Units (GRU).
Time (minutes)
THE MAVERICKTM ASSAY KIT
Maverick assay kits include an AutoArray, a Reagent Plate, and a USB drive.
The USB drive contains the assay protocol(s) and is linked to the
AutoArray and Reagent Plate via barcode. This enables the instrument
to track chip and reagent well usage allowing partial kits to be used
without danger of accidentally reusing a chip or well. It also prevents
incompatible kit components from being used.
Sample
Channel
1
2
Chip 1
1
9
17
2
10
18
3
11
19
4
12
20
5
13
21
6
14
22
7
15
23
8
16
24
The reagent plate is mapped out by chip
and the number of wells required to hold
the reagents specified by the protocol.
The reagent plate comes with all necessary
reagents preloaded into the appropriate wells.
Test samples are added to the numbered wells.
Each AutoArray contains 12 Maverick silicon photonic chips. Each of the 24 sample
channels (2 per chip) uses dedicated fluidics, providing the capacity the to test 24 samples.
Dedicated
fluidics for
each sample
1
2
3
4
5
6
7
8
9
11 13 15 17 19
21 23
10
12 14 16 18 20
22 24
EASY TO OPERATE
1. load sample
2. pipette to mix
3. insert usB, autoarray,
and reagent plate
into Maverick
4. press start.
instrument
automation moves
autoarray and
reagent plate as
directed by assay
protocol , testing up
to 24 samples with no
human interaction
APPLICATIONS
ISOTYPED IMMUNOGENICITY TESTING
MaverickTM Multi-Tier Anti-Drug Antibody Assay (MT-ADA)
Screen, Confirm and Isotype Anti-Drug Antibodies in One Assay
The Maverick™ MT-ADA assay streamlines immunogenicity testing by combining screening,
confirmation, and isotyping into a single assay providing real-time detection of
anti-drug antibodies (ADA) without the use of fluorescent probes or radioactive labels. The sensor spots on the assay chip are functionalized
with anti-isotype capture antibodies. This design
provides simultaneous ADA screening and
confirmation of eight classes/isotypes in the same
assay without additional effort or reagents.
The inherent multiplexing capability of the
Maverick also speeds assay development by
enabling assessment of assay sensitivity and
free drug tolerance for total ADA and
multiple classes/isotypes simultaneously.
Streptavidin
Coated Bead
Biotinylated
Drug
Sam
ple
Sa
mp
Cha
le
nne
Ch
l1
an
ne
l2
IgG3
IgG2b
IgG2a
IgG1
IgE
IgA
IgM
IgX
2
1
The isotype capture assay format allows both drug
binding sites on the ADA to be potentially available for
labeled drug binding. This enables the MT-ADA assay
to generate a signal with a single drug molecule bound
to the ADA as compared to a traditional bridging assay
which requires labeled drug to bind to both sites.
Anti-IgG1
Capture Ab
Maverick MT-ADA Sensorgram
1400
Igx
IgG1
1000
800
600
IgG3
400
IgG2a
IgM
200
IgG2b
IgE, IgA
0
-200
4
Running Buffer
6
8
10
12
Running Buffer
Time (minutes)
Running Buffer
Wash Buffer
2
Running Buffer
Genalyte Response Units (GRU)
1200
0
Sample
Confirmation w/ SA Beads
ADA (IgG1)
To demonstrate assay functionality,
mouse monoclonal antibodies of known
isotype against human insulin were used
to represent ADA and biotinylated insulin
was used to represent the drug.
Known concentrations of ADA were
spiked into a buffer sample and combined
with biotinylated drug using equilibrium
binding and acid dissociation methods.
Detection of ADAs with concentration
varying by isotype is demonstrated
in the sensorgram to the left.
MULTIPLEX ANTIBODY DETECTION IN MULTIPLE MATRICES
The Maverick performs assays in multiple complex matrices such as serum,
plasma, whole blood and crude extracts, minimizing sample preparation.
Serum
A multi-positive ENA serum sample tested on the Maverick ENA-7 assay kit diluted 1:50 in buffer.
Surfaces of individual sensor spots are functionalized with SLE-related autoantigens.
400
Antibody
Antigen
350
Genalyte Response Units (GRU)
Anti-IgG
Jo-1
Anti-Human IgG
(Pos Control)
SS-A60
SS-B
300
250
Scl-70
SS-A52
200
150
100
RNP
Smith
50
0
0.0
2.0
4.0
Time (minutes)
6.0
8.0
Cell Culture Media
Supernatant from a hybridoma culture producing mouse IgG1 monoclonal antibodies was tested
with a Maverick Mouse MT-ADA assay kit using an alternate protocol. Biotinylated rat αnti-mouse
kappa light chain antibody and streptavidin beads were used for signal amplification.
Streptavidin
Bead
Sensors
1400
Biotinylated Rat aMouse
Kappa Light Chain
1200
1000
IgG1 Mouse
Monoclonal Ab
Anti-IgG1
Capture Ab
800
600
400
200
0
0
2
4
Crude Extracts
Demonstration of the use of the Maverick to detect Bean Pod Mottle Virus (BPMV),
a common viral pathogen in soybean crops. The graph shows sensor response to
diluted (1:200) leaf extracts from BPMV infected and healthy soybean plants.
6
8
Time (minutes)
10
12
14
16
The sensor surface was functionalized
with anti-BPMV antibody.
Leaf extract samples were prepared by grinding
approximately 100 mg of leaf material in a mortar and
pestle with 1 mL of BSA–PBS buffer until homogeneous.
Solid plant debris was removed via centrifugation.
Source: M.S. McClellan et al.,
Biosensors and Bioelectronics 31 (2012) 388-392
RAPID ASSAY DEVELOPMENT
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I13
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I14
d
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Use
PI
Not
Capture Ab
De
All pair-wise combinations of 15
insulin antibodies were evaluated as
both capture antibody (spotted on chip)
and biotinylated detection antibody (in
reagent plate). The analyte (insulin), buffer
conditions, and secondary (streptavidin
coated beads) were the same for all assays.
te
c
An tion
al A
yt b
Bu e
ffe
Be r
ad
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se
co
nd
ar
y
)
The Maverick’s onboard AutoArray and Reagent Plate automation enable rapid
testing of multiple assay conditions in a single run, streamlining assay development.
The contents of each well will pass over all sensors in the corresponding sample channel of its
designated chip, allowing multiple combinations to be tested at once. The design of the Maverick AutoArray
(12 chips x 2 sample channels x 16 sensor spots) enables an assay developer to test up to 24 different
analytes and/or 24 different buffer conditions against up to 16 different probe molecules in a single run.
Genalyte’s custom spotting service can economically functionalize assay chips with client probe molecules
and provide fully assembled AutoArrays. The open reagent plate format allows a user to load reagents of
their choice into the reagent wells. Guidance is provided by a plate map automatically generated by the
Maverick Protocol Editor software based on the assay protocol designed by the user.
Screening of Antibody Pairs in Development of a Quantitative Insulin Assay
I7
The top candidate pairs (yellow in
table, left) clearly stand out on the
real-time sensorgrams (lower right).
Dose curves for the top two
candidate pairs lead to a clear
winning combination (lower left).
Real-Time Sensorgrams for Top 2 Pairs
DESIGN YOUR OWN PROTOCOLS
Maverick Protocol Editor software simplifies assay development by enabling researchers to design their own
protocol or edit an existing protocol for any Maverick assay kit. The software generates a reagent plate map
based on the protocol to guide the user in loading reagents.
Create/modify reagent list
Reagent list populates
Specify reagent flow Software calculates
drop down menu to add rate and duration
required well volume
additional protocol steps
DIMENSIONS
Length 17.75 in / 45 cm
Width 15 in / 38 cm
Height 21 in / 53 cm
WEIGHT
87 lb / 39 kg
POWER REQUIREMENTS
110/220 volt compatible
SCALABLE
Link systems for
increased throughput
ORDERING INFORMATION
85000
Maverick M24 Detection System
85001
Maverick M24 XM Expansion Unit
To place an order please call
858-956-1200
[email protected]
Genalyte
11095 Flintkote Ave Suite A
San Diego, CA 92121
Research Use Only
Genalyte, Inc. | 858.956.1200 | [email protected] | www.genalyte.com
Maverick is a trademark of Genalyte | © 2013 Genalyte
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