Clinical mass spectrometry test for apolipoprotein C-III proteoforms

PROJECT SUMMARY Only about dozen mass spectrometryMSprotein tests have been translated into clinical laboratories since the MALDI and ESI approaches were developed thirty years agoWhile the cost and complexity of these tests are important factors impeding their clinical adoptionnew content generated via proteoforms detection could provide the impetus for further development and translationOur recent studies have revealed strong associations between relative abundance of a specific apolipoprotein C III proteoforms and plasma triglycerides concentrations in multiple cross sectional and longitudinal cohortsand also suggested prognostic and possible mechanistic role in dyslipidemia and cardiovascular disease riskThese intriguing results warrant further large scale investigations to verify the biomarker potential of these proteoforms in lipid metabolism and cardiometabolic riskWe propose to develop and demonstrate a low costhigh throughput MALDI TOF MS apoC III proteoforms test that will find immediate use in clinical utility studies of these proteoformsThe test workflow will include immunoaffinity isolaton of apoC III from minimal amounts of human plasma with antibody coatedwell platesand elution onto MALDI targets for MS analyses and determination of the proteforms relative abundanceThe steps of antibody immobilizationsample dilutionassayingrinsesand elution onto MALDI targets will be optimizedThe number of assay steps will be kept to a minimumto keep the cost per test lowReproducibility of the new test will also be evaluatedThe proposed MS test is innovative in its simplicitycostand content deliveredit will transform clinical research and diagnostics by enabling fast and cost effective screening of apoC III proteoformsSimilar tests for other proteins can also be developedsignificantly expanding our ability to study human proteoforms and their role in disease diagnosistherapy monitoringand outcome PROJECT NARRATIVE The new apolipoprotein C III MS test will transform clinical research by enabling fast and cost effective screening of apoC III proteoformsThe new test could find rapid use in extensive clinical studies of the proteoforms that will expand our understanding of apoC pathophysiology and its impact in lipid metabolism and cardiometabolic risk