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Principal Investigator: Wei-Jun Qian
Organization: BATTELLE PACIFIC NORTHWEST LABORATORIES
Fiscal Year: 2024
Award: $949,078
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
Project Summary/Abstract:
Type 1 diabetes (T1D) is a devastating disease often occurring in children and young adults resulting from the
autoimmune-mediated loss of pancreatic β-cells. It has been challenging for monitoring the disease progression
and the efficacy of clinical interventions. Therefore, a critical need remains for highly reliable assays that quantify
proteins or peptide hormones (e.g., insulin, glucagon) and their specific isoforms (or proteoforms) as markers of
endocrine and exocrine function. Such assays will play an important role in facilitating effective monitoring of
disease progression or efficacy of novel clinical interventions prior to or following the onset of T1D. Most current
clinical assays depend on the use of antibodies or other affinity reagents almost exclusively. However, the exact
specificity of affinity reagents is often unknown or difficult to characterize. Targeted mass spectrometry (MS)
presents a promising alternative to immunoassays. Therefore, the overall objective of this application is to
develop reliable, proteoform-specific, and multiplex targeted MS assays for a list of protein/peptide analytes of
significance in T1D. Specifically, we aim to develop multiplex targeted MS assays for the following panel of
targets as markers of endocrine and exocrine function: insulin, glucagon, amylin, chromogranin, somatostatin,
prohormone isoforms (e.g., proinsulin, proglucagon), hybrid insulin peptides, trypsinogen, glycated CD59, as
well as other markers of interest to the T1D research community. To facilitate full validation of the robustness
and transferability of the assays, the overall objective will be accomplished through the collaborative efforts of
two independent targeted MS labs and through a multi-lab assay validation effort. Specifically, Aim 1 will be
focused on establishing optimal assay configurations for confident detection of endogenous analytes in serum
samples for specific proteoforms or peptides of interest in T1D. Aim 2 will be centered on assay optimization and
full assay characterization in the aspects of reproducibility, stability, selectivity, linearity, and limit of
quantification. Inter-lab assay protocol transfer and assay characterization will also be pursued. Aim 3 will
demonstrate the robustness and utility of the assays through multi-lab validation of the assays by analyzing the
same cohort of clinical serum samples and benchmarking against well-established immunoassays for selected
analytes such as insulin and c-peptide. Together, the project will establish highly reliable and easy-to-transfer
multiplex targeted MS assays for many difficult to measure T1D markers. These assays are expected to make
a significant contribution to the monitoring of pancreatic endocrine/exocrine functions, the disease progression,
as well as the efficacy of clinical interventions in T1D research.
Terms: <0-11 years old><Affinity><Antibodies><Antidiabetic Hormone><Assay><Autoantibodies><Autoimmune><B9 endocrine pancreas><Benchmarking><Best Practice Analysis><Bioassay><Biological Assay><Blinded><Blood Glucose><Blood Serum><Blood Sugar><Brittle Diabetes Mellitus><C-Peptide><Chemical Fractionation><Child><Child Youth><Children (0-21)><Children's Hospital><Chromogranins><Clinical><Clinical Chemistry><Clinical Research><Clinical Study><Collaborations><Communities><Coupled><Cyclic Somatostatin><Detection><Disease><Disease Progression><Disorder><Endocrine><Endocrine Pancreas><FRACN><Failure><Fractionation><Fractionation Radiotherapy><Genes><Glucagon><Glukagon><Goals><Growth Hormone Inhibiting Factors><Growth Hormone-Inhibiting Hormone><HG-Factor><High Throughput Assay><Human><Humulin R><Hybrids><Hyperglycemic-Glycogenolytic Factor><IDDM><Immunoassay><Indiana><Insulin><Insulin-Dependent Diabetes Mellitus><Insulinoma amyloid peptide><Intervention><Intervention Strategies><Islands of Langerhans><Islets of Langerhans><Isoforms><Juvenile-Onset Diabetes Mellitus><Ketosis-Prone Diabetes Mellitus><Laboratories><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measures><Mediating><Medicine><Methods><Modern Man><Monitor><Nesidioblasts><Novolin R><Pancreatic Islets><Pancreatic beta Cell><Pancreatic β-Cell><Pars endocrina pancreatis><Pathogenesis><Patients><Pediatric Hospitals><Peptide Hormone Gene><Peptides><Performance><Philadelphia><Play><Post-Translational Modification Protein/Amino Acid Biochemistry><Post-Translational Modifications><Post-Translational Protein Modification><Post-Translational Protein Processing><Posttranslational Modifications><Posttranslational Protein Processing><Proinsulin><Protein Isoforms><Protein Modification><Proteins><Protocol><Protocols documentation><Publications><RNA Splicing><Reagent><Regular Insulin><Reporting><Reproducibility><Research><Resolution><Role><SRIH><SRIH-14><Sampling><Scientific Publication><Serum><Somatostatin><Somatostatin-14><Somatotropin Release Inhibiting Factors><Somatotropin Release-Inhibiting Hormone><Specificity><Splicing><Standardization><Structure of beta Cell of islet><Sudden-Onset Diabetes Mellitus><T1 DM><T1 diabetes><T1D><T1DM><Trypsinogen><Type 1 Diabetes Mellitus><Type 1 diabetes><Type I Diabetes Mellitus><Universities><Validation><Variant><Variation><Work><adult youth><amlintide><amylin><assay development><autoimmune antibody><autoreactive antibody><benchmark><clinical applicability><clinical application><clinical efficacy><cohort><connecting peptide><diabetes associated peptide><diabetes pathogenesis><experience><experiment><experimental research><experimental study><experiments><glycation><growth hormone release inhibiting factor><high throughput screening><insulin dependent diabetes><insulin dependent diabetes mellitus onset><insulin dependent type 1><insulinoma amyloid polypeptide><interest><interventional strategy><islet amyloid polypeptide><islet progenitor><juvenile diabetes><juvenile diabetes mellitus><ketosis prone diabetes><kids><multiplex assay><nano><nanobodies><nanobody><non-enzymatic glycosylation><nonenzymatic glycosylation><novel><pancreas beta cell><pancreas β cell><pancreatic amylin><pancreatic b-cell><peptide hormone><pressure><pro-glucagon><proglucagon><prohormone><resolutions><sdAb><self reactive antibody><serological marker><single domain antibodies><social role><type 1 diabetes onset><type I diabetes><type one diabetes><validation studies><validations><young adult><young adulthood><youngster>