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Middle-throughput LC-MS-based platelet proteomics with minute sample amounts using semi-automated positive pressure FASP in 384-well format (PF384).
Thromb Haemost
January 2025
Department of Bioinformatics, Biocenter, University of Würzburg, Wurzburg, Germany.
Comprehensive characterization of platelets requires various functional assays and analysis techniques, including omics-disciplines, each requiring an individual aliquot of a given sample. Consequently, the sample material per assay is often highly limited rendering downscaling a prerequisite for effective sample exploitation. Here we present a transfer of our recently introduced 96-well-based proteomics workflow (PF96) into the 384-well format (PF384) allowing for a significant increase in sensitivity when processing minute platelet protein amounts.
View Article and Find Full Text PDFAutomated high-throughput RP-HPLC-MS and SFC-MS analytical and purification platforms to support drug discovery.
J Chromatogr A
December 2024
Chemistry Capabilities, Analytical & Purification, Global Discovery Chemistry. Janssen Research & Development, a Division of Janssen Pharmaceuticals, Johnson & Johnson company, 1400 McKean Rd. Spring House PA 19477, USA.
In recent years, the need to accelerate drug discovery processes in the pharmaceutical industry has revived the interest of implementing automated workflows, allowing the simultaneous processing of multiple samples on global processes that are referred as High-Throughput Purification (HTP). In this work, SAPIO Laboratory Information Management System (SAPIO LIMS) has been customized at the HTP laboratories of Janssen R&D to accommodate the needs of global purification groups on several automated HTP workflows, integrating Analytical Studio™ data processing tool on multiple steps. Herein we describe the workflow details from crude analysis via RP-LC-MS or SFC-MS systems to sample redissolution and delivery to Compound Logistics (CL) in tubes ready for assay plate preparation.
View Article and Find Full Text PDFPlasma S100β is a predictor for pathology and cognitive decline in Alzheimer's disease.
Fluids Barriers CNS
January 2025
Sanders-Brown Center on Aging, College of Medicine, University of Kentucky, 760 Press Ave, 124 HKRB, Lexington, KY, 40536-0679, USA.
Background: Blood-brain barrier dysfunction is one characteristic of Alzheimer's disease (AD) and is recognized as both a cause and consequence of the pathological cascade leading to cognitive decline. The goal of this study was to assess markers for barrier dysfunction in postmortem tissue samples from research participants who were either cognitively normal individuals (CNI) or diagnosed with AD at the time of autopsy and determine to what extent these markers are associated with AD neuropathologic changes (ADNC) and cognitive impairment.
Methods: We used postmortem brain tissue and plasma samples from 19 participants: 9 CNI and 10 AD dementia patients who had come to autopsy from the University of Kentucky AD Research Center (UK-ADRC) community-based cohort; all cases with dementia had confirmed severe ADNC.
High-throughput determination of exchange rates of unmodified and PTM-containing peptides using HX-MS.
Mol Cell Proteomics
January 2025
Broad Institute of MIT & Harvard, Cambridge, MA. Electronic address:
Despite the widespread use of MS for hydrogen/deuterium exchange measurements, no systematic, large-scale study has been conducted to compare the observed exchange rates in protein-derived, unstructured peptides measured by MS to the predicted exchange rates calculated from NMR-derived values and how neighboring residues and post-translational modifications influence those exchange rates. In this study, we sought to test the accuracy of predicted values by performing hydrogen exchange measurements on whole cell digests to generate an unbiased dataset of 563 unique peptides derived from naturally-occurring protein sequences. A remarkable 97% of observed exchange rates of peptides are within two-fold of predicted values.
View Article and Find Full Text PDFMicrovascular Metrics on Diabetic Retinopathy Severity: Analysis of Diabetic Eye Images from Real-World Data.
Biomedicines
December 2024
BCN Peptides, S.A., Polígono Industrial Els Vinyets-Els Fogars II, Sant Quintí de Mediona, 08777 Barcelona, Spain.
To quantify microvascular lesions in a large real-world data (RWD) set, based on single central retinal fundus images of diabetic eyes from different origins, with the aim of validating its use as a precision tool for classifying diabetic retinopathy (DR) severity. Retrospective meta-analysis across multiple fundus image datasets. The study analyzed 2445 retinal fundus images from diabetic patients across four diverse RWD international datasets, including populations from Spain, India, China and the US.
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