SAMPLE PREPARATION; The Achilles Heel of Mass Spectrometry Based Diagnostics II.

SAMPLE PREPARATIONThe Achilles Heel of Mass Spectrometry Based Diagnostics IIProject SummaryLC MS provides superior data in clinical diagnostics relative to immunoassaysbut it is slow and costlyAs the old saying goestime is moneyThe technique must become simplerfasterand less expensive to displace olderless accurate immunological assayIAmethodsThe focus of this proposal is to make LC MS analytics more competitive in clinical diagnostics by increasing throughputfold with existing instrumentationThe objective in Phase I was to use analyte sequestering transport particlesASTPsto circumvent the above limitations by istructure specifically sequestering analytes of interestiiseparating them from non analytes within a miniirapidly transferring them to an MSand ivenabling the MS to identify all analytes of interest in less thansecThis was achieved by preparingmegadaltonmDaASTPs that sequester analytes at their surfaceelevating their effective Mw tomDa and allowing them to be purified by mobile affinity sorbent chromatographyMASCinsecThermal or solvent dissociation was used to recover analytes for ESI MS or LDTD MS wellsThis proposal has four specific aimsOne is to complete manufacturing protocols for antibody and protein A G based ASTP products that captureresolveand transport analytes to MS or fluorescence instruments in researchbig pharmaand CRO laboratoriesThe second is to integrate the above ASTP products into miniature membrane laboratoriesMemLabsproducing point of collection products that extract plasma from a drop of blood and prepare samples in situ for biomarker detectionThe third aim was to evaluate and optimize the release and detection of analytes from ASTPs via LDTD MSMALDI MSand ESI MSThe fourth specific aim is to adapt ASTP technology to multiplexed drug testing and the analysis of protein iprimary structureiipost translational modificationsand iiiconformation PROJECT NARRATIVEDelivery of high quality medicine without bankrupting the economy is of major importance todaymass spectrometryMSbased assays being a good exampleAlthough MS is inherently high throughputsample preparation is slow and costlyThe significance of this proposal is in making high quality MS based immunoassays widely available to the life science world by eliminating conventional sample preparation laboratories and reducing analysis costsXThis will be achieved by advances in sample collectionpreparationand throughput that circumvent conventional mode of preparing samples for LC MSamong the more innovative beinga miniature membrane laboratory bearingnanoparticulate analyte sequestering transport particles that select clusters of targeted biomarkers and rapidly transport them throughmobile sorbent affinity chromatographyMASCto an MSThe net effect of implementing these technologies is that superior MS assay methods will be costs competitive with conventional immunoassays