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Robust Microflow LC-MS/MS for Proteome Analysis: 38 000 Runs and Counting.
Bian, Yangyang; Bayer, Florian P; Chang, Yun-Chien; Meng, Chen; Hoefer, Stefanie; Deng, Nan; Zheng, Runsheng; Boychenko, Oleksandr; Kuster, Bernhard.
Afiliación
  • Bian Y; Chair of Proteomics and Bioanalytics, Technical University of Munich, Emil Erlenmeyer Forum 5, 85354 Freising, Germany.
  • Bayer FP; College of Life Science, Northwest University, Taibai North Road 229, Xi'an, Shaanxi 710069, P.R. China.
  • Chang YC; Chair of Proteomics and Bioanalytics, Technical University of Munich, Emil Erlenmeyer Forum 5, 85354 Freising, Germany.
  • Meng C; Chair of Proteomics and Bioanalytics, Technical University of Munich, Emil Erlenmeyer Forum 5, 85354 Freising, Germany.
  • Hoefer S; Bavarian Biomolecular Mass Spectrometry Center (BayBioMS), Technical University of Munich, Gregor-Mendel-Straße 4, 85354 Freising, Germany.
  • Deng N; Chair of Proteomics and Bioanalytics, Technical University of Munich, Emil Erlenmeyer Forum 5, 85354 Freising, Germany.
  • Zheng R; Instrumental Analysis Center, Xi'an Jiaotong University, No. 28, Xianning West Road, Xi'an, Shaanxi 710049, P.R. China.
  • Boychenko O; Thermo Fisher Scientific, Dornierstraße 4, 82110 Germering, Germany.
  • Kuster B; Thermo Fisher Scientific, Dornierstraße 4, 82110 Germering, Germany.
Anal Chem ; 93(8): 3686-3690, 2021 03 02.
Article en En | MEDLINE | ID: mdl-33596053
Microflow liquid chromatography tandem mass spectrometry (µLC-MS/MS) is becoming a viable alternative to nanoflow LC-MS/MS for the analysis of proteomes. We have recently demonstrated the potential of such a system operating with a 1 mm i.d. × 150 mm column and at a flow rate of 50 µL/min for high-throughput applications. On the basis of the analysis of ∼38 000 samples measured on two instruments over the past two years, we now show that the approach is extremely robust. Up to 1500 analyses were performed within one month, and >14 000 samples could be analyzed on a single column without loss of chromatographic performance. Samples included proteomes of cell lines, tissues, and human body fluids, which were analyzed with or without prior peptide fractionation or stable isotope labeling. We show that the µLC-MS/MS system is capable of measuring 2600 proteins from undepleted human plasma and ∼5000 proteins from crude human urine in 1 day, demonstrating its potential for in-depth as well as high-throughput clinical application.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteoma / Espectrometría de Masas en Tándem Límite: Humans Idioma: En Revista: Anal Chem Año: 2021 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteoma / Espectrometría de Masas en Tándem Límite: Humans Idioma: En Revista: Anal Chem Año: 2021 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos