msd technical brief gtp meso scale discovery … · rock rho fak crk pax pyk2 gtp g˜ -o gefs cdc42...
TRANSCRIPT
MSD -Meso Scale Discovery
Applications include:
Signal transduction path way evaluation
ROCK
Rho
FAK CrK
Pax PYK2
GTP Gα-O
GEFs
CDC42 Rac
MEKs
PAK MKKs
ASK1
JNK
P38 IP3R
PKC
BIMP1
BCL10
NFκB IκBs
P
IKKs
IκBsDegradation
Src
SOSGRB2SHC
RasGEFs
Raf1
MEKs
ERKs
PLC-β
DAG
NFκB
IKKs
PκB
IP3
Ca+2Ca+2
Ca+2
Calm
P
Caln
NFAT
NFAT
P
PI3K
Gβ Gγ
LARG
BRaf
MEKs
ERKs
cAMP
EPACPKA
PDZRhoGEF
Gα-1 GTP
Rap1
p115RhoGEF
RT
KS
CREB
Gα12/13
SRF
GTP
CREB
SRF ELK-1
c-Jun
c-Fos
STAT
NFκBNFAT
Gene Expression
Gα Gβ
© 2013 CDT all rights reserved
CON
LFU
ENCE
DIS
COVE
RY T
ECH
NO
LOG
IES
| TE
CHN
ICAL
BRI
EFTECHNICAL BRIEF:
MSD -Meso Scale DiscoveryD I S C O V E R Y T E C H N O L O G I E S
The Meso Scale Discovery (MSD) platform provides a faster alternative to traditional ELISAs for performing sand-wich immunoassays. The MSD assay platform utilizes Ruthenium (II) tris-bipyridine-(4-methylsulfone) [Ru(bpy)3] that, once conjugated to the analyte, serves as the tracer in competitive assays. The Ru(bpy)3-based tag undergoes a rapid redox reaction that emits light in the presence of an applied voltage. Assay kits for several eicosanoids have been adapted to the MSD platform. The Confluence Discovery Technologies (CDT) team has access to dozens of kits avail-able across therapeutic areas. An optimized MSD electro-chemiluminescent assay can quantitate host cell proteins with a simple protocol few wash steps and low cost. The MSD assay offers several technical advantages as an assay platform:
•Capture: 10-100 fold increase compared to ELISA
•Dynamic Range: 3-4 log vs. 1-1.5 log range for ELISA
•Sensitivity: Detection of analytes at sub-picogram levels
•Reduced Sample Volume: 5-25 µl volume vs. 50-100 µl for ELISA
•Speed: The entire assay can be performed in 3 hours
•Multiplex Capability: Simultaneous measurements for up to 10 analytes
Applications
The CDT team has a wide range of experience in develop-ing and validating MSD assays to suit many different requirements. Our highly trained and knowledgeable scien-tists will assist with experimental design to develop and vali-date assays to meet client-specified criteria. We also perform data analysis and interpretation of MSD data to enable our clients to obtain the best insights from large, complex data outputs.
ROCK
Rho
FAK CrK
Pax PYK2
GTP Gα-O
GEFs
CDC42 Rac
MEKs
PAK MKKs
ASK1
JNK
P38 IP3R
PKC
BIMP1
BCL10
NFκB IκBs
P
IKKs
IκBsDegradation
Src
SOSGRB2SHC
RasGEFs
Raf1
MEKs
ERKs
PLC-β
DAG
NFκB
IKKs
PκB
IP3
Ca+2Ca+2
Ca+2
Calm
P
Caln
NFAT
NFAT
P
PI3K
Gβ Gγ
LARG
BRaf
MEKs
ERKs
cAMP
EPACPKA
PDZRhoGEF
Gα-1 GTP
Rap1
p115RhoGEF
RT
KS
CREB
Gα12/13
SRF
GTP
CREB
SRF ELK-1
c-Jun
c-Fos
STAT
NFκBNFAT
Gene Expression
Gα Gβ
© 2013 CDT all rights reserved
CDTv1.8.13
Confluence Discovery Technologies, Inc.4320 Forest Park Avenue
Saint Louis, MO 63108Phone: 314-932-4032 x305
Fax: 314-932-4038www.confluencediscovery.com
D I S C O V E R Y T E C H N O L O G I E S
CAPABILITIESD I S C O V E R Y T E C H N O L O G I E S
CON
LFU
ENCE
DIS
COVE
RY T
ECH
NO
LOG
IES
| CA
PABI
LITI
ES
Leaders in biochemistry and mechanistic enzymology, our senior team has access to a suite of capabilities and technologies enabling: • Drug:Target interaction • Enzyme:Inhibitor mechanisms • Direct binding kinetics • Compound mechanism of action
Con�uence o�ers a spectrum of cell and molecular biology capabilities: • Identi�cation of biologically important signal transduction pathways • Quantitation of e�cacy and selectivity • Understanding safety issues linked to pathways • De-convolution to isolate pathway nodes • Evaluation of multiple targets within the pathway • Identi�cation of key pathway point(s) for therapeutic intervention • Target validation, cloning, expression, and puri�cation • Custom assay development • High-throughput screening
Our team delivers when “OFF the SHELF” is not an option: • Assay development, optimization, and validation • Multiplex technology platforms and instrumentation • Complex data analysis • Specialty screens for preclinical technical application • Trouble shooting and problem solving
Our team provides clients with the expertise to: • Identify target, mechanism, and disease biomarkers • Select the appropriate biomarkers for a drug discovery program • Execute biomarker assay design, development, validation, and interpretation for pre-clinical and clinical studies • Seamless assay transfer from pre-clinical to clinical studies
Our medicinal and computational chemists are experts in designing and delivering novel chemical entities through: • Schrödinger® enabled computational drug design • High throughput hit prioritization • Chemical series prioritization • Structure activity relationship generation • Custom synthesis