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Meso Scale Discovery and Luminex Comparative Analysis of Calbindin D28K 外文参考文献
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HindawiPublishingCorporation
JournalofBiomedicineandBiotechnology
Volume2009,ArticleIDpages
doi:10.7426
ResearchArticle
MesoScaleDiscoveryandLuminexComparativeAnalysisof
CalbindinD28K
SamerSourial,MarithaMarcusson-St?ahl,andKarinCederbrant
MolecularToxicology,SafetyAssessment,AstraZenecaR&DS¨odert¨alje,15185S¨odert¨alje,Sweden
CorrespondenceshouldbeaddressedtoSamerSourial,s.sourial@student.liverpool.ac.uk
Received6May2009;Accepted23July2009
RecommendedbyColinCooper
Thesensitivityofdi?erentrenalregionstoxenobioticsrequiresthedevelopmentofamultipleximmunoassayforthesimultaneous
analysisofkidneybiomarkers.CalbindinD28Kisadistaltubule-speci?cproteinthatcanbedetectedinurineunderpathological
conditions.Inthisstudy,apairofanti-parativeanalysisoftheimmunoassaywasperformedontheMeso
ScaleDevelopment(MSD)andLuminexplatforms.AnalysisonbothplatformsdetectedcalbindinD28Kconcentrationsbetween
100ng/mLand100pg/mL.Luminexdetected10-foldtheamountofcalbindinD28KinsamplesanalyzedascomparedtoMSD,
whereas calbindin D28K level in rat and human urine was below detection limit in both platforms. The application of the
immunoassaysdescribedhereinmaybeusefulintoxicologicalandpathologicalstudiesofdistaltubulardamageinratsandhuman.
Copyright?2009SamerSourialetal.ThisisanopenaccessarticledistributedundertheCreativeCommonsAttributionLicense,
whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperlycited.
1.Introduction
speci?c damage before functional loss. Limited sample
volume and excess costs have led to the development of
multiplex immunoassays for the simultaneous analysis of
multiplebiomarkers.
The high renal blood ?ow, renal biotransformation of
chemicalstoreactivemetabolites,andthenephronsability
to concentrate tubular ?uid render the kidney sensitive
to xenobiotics. Due to the kidneys regional sensitivit
正在加载中，请稍后...Meso&Scale&Discovery
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Walkthrough
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Meso Scale Discovery's technology
uses proprietary MULTI-ARRAY& and MULTI-SPOT&
microplates with electrodes integrated into the bottom of the
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MSD's electrodes are made from
carbon, an excellent material for biological assays. Biological
reagents can be attached to the carbon simply by passive adsorption
and retain a high level of biological activity.
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MSD assays use
electrochemi-luminescent labels for ultra-sensitive detection.
These labels are non-radioactive, stable and offer a choice of
convenient coupling chemistries.
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TITLE="Meso&Scale&Discovery" />
Electrochemiluminescent labels emit
light when electrochemically stimulated. The detection process is
initiated at electrodes located in the bottom of MSD's microplates.
Only labels near the electrode are excited and detected, enabling
non-washed assays.
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TITLE="Meso&Scale&Discovery" />
MSD Read Buffers contain coreactants
that enhance the electrochemi-luminescence signals. These
coreactants are also stimulated when in proximity to the electrodes
in the microplate.
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TITLE="Meso&Scale&Discovery" />
MSD's labels emit light at 620 nm,
eliminating problems with color quenching. Few compounds interfere
with the electrochemiluinescence process so you can use large,
diverse libraries with confidence.
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Multiple excitation cycles of each
label amplify the signal to enhance light levels and improve
sensitivity. Background signals are minimal because the stimulation
mechanism (electricity) is decoupled from the signal (light).
已投稿到：Utilization of Meso Scale Discovery ELISA technology to quantitate the in vivo pharmacodynamic effects of Rapamycin treatment on the AKT pathway | Cancer Research
Utilization of Meso Scale Discovery ELISA technology to quantitate the in vivo pharmacodynamic effects of Rapamycin treatment on the AKT pathway
Lillian Sams, George Rodgers, Laura Nickell, Tao Wang, Rose Ajamie, Everett Perkins and Sandaruwan Geeganage
Lillian SamsLilly Research Laboratories, Indianapolis, INGeorge RodgersLilly Research Laboratories, Indianapolis, INLaura NickellLilly Research Laboratories, Indianapolis, INTao WangLilly Research Laboratories, Indianapolis, INRose AjamieLilly Research Laboratories, Indianapolis, INEverett PerkinsLilly Research Laboratories, Indianapolis, INSandaruwan GeeganageLilly Research Laboratories, Indianapolis, IN
AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CAAbstract
The integral role of multiple members of the PI3K/AKT pathway in cell survival has made this pathway a fertile hunting ground for the development of targeted cancer therapeutics. Generation of an optimal therapeutic agent to a specific target requires thorough characterization. This characterization often involves repetitive assessment of the effects of hundreds of related structures not only on the direct downstream effectors of the target, but on other members of the pathway of interest as well. The collection of in vitro cell culture data is valuable for the initial assessment of compound related target effects. Ultimately however, it is essential to evaluate the effects of treatment on members of the pathway and to directly demonstrate that the target is inhibited in vivo. Quantitating the phosphorylation status of multiple signaling pathway members and downstream effectors in treated tumor samples via western blot analysis is laborious and time-consuming, particularly in a medium throughput mode. Using the Meso Scale Discovery (MSD) ELISA technology, we were able to rapidly and efficiently assess the in vivo effects of treatment with the mTOR inhibitor Rapamycin on multiple phosphorylation sites for AKT, p70S6 Kinase, S6RP and GSK3β in HCT 116 xenograft tumors. Utilization of the MSD technology not only facilitated the thorough in vivo characterization of Rapamycin treatment on several members of the PI3K/AKT pathway, but allows for enhanced compound throughput without diminution of data collection during the course of an SAR effort for the development of a targeted cancer therapeutic agent.Footnotes98th AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA
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Lillian Sams, George Rodgers, Laura Nickell, Tao Wang, Rose Ajamie, Everett Perkins and Sandaruwan Geeganage
Cancer Res (67) (9 Supplement) 2830;
Citation Manager Formats
Utilization of Meso Scale Discovery ELISA technology to quantitate the in vivo pharmacodynamic effects of Rapamycin treatment on the AKT pathway
Lillian Sams, George Rodgers, Laura Nickell, Tao Wang, Rose Ajamie, Everett Perkins and Sandaruwan Geeganage
Cancer Res (67) (9 Supplement) 2830;
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