Tryptic digestion of human serum for proteomic mass spectrometry automated by centrifugal microfluidics

Lab on a Chip ◽  
2020 ◽  
Vol 20 (16) ◽  
pp. 2937-2946
Author(s):  
J.-N. Klatt ◽  
M. Depke ◽  
N. Goswami ◽  
N. Paust ◽  
R. Zengerle ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Human Serum ◽  
Centrifugal Microfluidics ◽  
Tryptic Digestion

Tryptic digestion of human serum automated by centrifugal microfluidics.

scholarly journals Oxidized plant sterols in human serum and lipid infusions as measured by combined gas-liquid chromatography-mass spectrometry

2001 ◽  
Vol 42 (12) ◽  
pp. 2030-2038 ◽  
Author(s):  
Jogchum Plat ◽  
Harald Brzezinka ◽  
Dieter Lütjohann ◽  
Ronald P. Mensink ◽  
Klaus von Bergmann
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Human Serum ◽  
Plant Sterols ◽  
Gas Liquid Chromatography ◽  
Liquid Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry

scholarly journals Protein Turnover Measurements in Human Serum by Serial Immunoaffinity LC-MS/MS

2018 ◽  
Vol 64 (2) ◽  
pp. 279-288 ◽  
Author(s):  
Vahid Farrokhi ◽  
Xiaoying Chen ◽  
Hendrik Neubert
Keyword(s):  
Mass Spectrometry ◽  
Cell Adhesion ◽  
Human Serum ◽  
Adhesion Molecule ◽  
Protein Turnover ◽  
Cell Adhesion Molecule ◽  
Analytical Sensitivity ◽  
Target Proteins ◽  
Cell Adhesion Molecule 1 ◽  
Sample Set

Abstract BACKGROUND The half-life of target proteins is frequently an important parameter in mechanistic pharmacokinetic and pharmacodynamic (PK/PD) modeling of biotherapeutics. Clinical studies for accurate measurement of physiologically relevant protein turnover can reduce the uncertainty in PK/PD model-based predictions, for example, of the therapeutic dose and dosing regimen in first-in-human clinical trials. METHODS We used a targeted mass spectrometry work flow based on serial immunoaffinity enrichment ofmultiple human serum proteins from a [5,5,5-2H3]-L-leucine tracer pulse-chase study in healthy volunteers. To confirm the reproducibility of turnover measurements from serial immunoaffinity enrichment, multiple aliquots from the same sample set were subjected to protein turnover analysis in varying order. Tracer incorporation was measured by multiple–reaction-monitoring mass spectrometry and target turnover was calculated using a four-compartment pharmacokinetic model. RESULTS Five proteins of clinical or therapeutic relevance including soluble tumor necrosis factor receptor superfamily member 12A, tissue factor pathway inhibitor, soluble interleukin 1 receptor like 1, soluble mucosal addressin cell adhesion molecule 1, and muscle-specific creatine kinase were sequentially subjected to turnover analysis from the same human serum sample. Calculated half-lives ranged from 5–15 h; however, no tracer incorporation was observed for mucosal addressin cell adhesion molecule 1. CONCLUSIONS The utility of clinical pulse-chase studies to investigate protein turnover can be extended by serial immunoaffinity enrichment of target proteins. Turnover analysis from serum and subsequently from remaining supernatants provided analytical sensitivity and reproducibility for multiple human target proteins in the same sample set, irrespective of the order of analysis.

scholarly journals Multiple Heart-Cutting Two Dimensional Liquid Chromatography And Isotope Dilution Tandem Mass Spectrometry For The Absolute Quantification Of Proteins In Human Serum

2021 ◽  
pp. 339022
Author(s):  
Amanda Suárez Fernández ◽  
Pablo Rodríguez-González ◽  
Lydia Álvarez ◽  
Montserrat García ◽  
Héctor González Iglesias ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Human Serum ◽  
Isotope Dilution ◽  
Absolute Quantification ◽  
Tandem Mass ◽  
Two Dimensional ◽  
The Absolute

scholarly journals Analysis of Endosulfan and Its Metabolites in Human Serum Using Gas Chromatography-Tandem Mass Spectrometry

2001 ◽  
Vol 39 (5) ◽  
pp. 177-182 ◽  
Author(s):  
F. J. Arrebola ◽  
J. L. Martinez Vidal ◽  
A. Fernandez-Gutierrez
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Tandem Mass Spectrometry ◽  
Human Serum ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry
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