TY - JOUR A1 - Müller, Larissa A1 - Techritz, Sandra A1 - Jakubowski, Norbert A1 - Roos, P.H. T1 - A multi-parametric microarray for protein profiling: simultaneous analysis of 8 different cytochromes via differentially element tagged antibodies and laser ablation ICP-MS N2 - The paper presents a new multi-parametric protein microarray embracing the multi-analyte capabilities of laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). The combination of high throughput reverse phase protein microarrays with element tagged antibodies and LA-ICP-MS makes it possible to detect and quantify many proteins or biomarkers in multiple samples simultaneously. A proof of concept experiment is performed for the analysis of cytochromes particularly of cytochrome P450 enzymes, which play an important role in the metabolism of xenobiotics such as toxicants and drugs. With the aid of the LA-ICP-MS based multi-parametric reverse phase protein microarray it was possible to analyse 8 cytochromes in 14 different proteomes in one run. The methodology shows excellent detection limits in the lower amol range and a very good linearity of R² ≥ 0.9996 which is a prerequisite for the development of further quantification strategies. KW - Multi-parametric KW - Multiplexing KW - Microarray KW - Immunoassay KW - LA-ICP-MS KW - Cytochrome P450 PY - 2013 DO - https://doi.org/10.1039/c3an00468f SN - 0003-2654 SN - 1364-5528 VL - 138 IS - 21 SP - 6309 EP - 6315 PB - Royal Society of Chemistry CY - Cambridge AN - OPUS4-29275 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - The Role of Modern Plasma-based Spectroscopies for Development and Characterization of Reference Materials T2 - Winter Conference on Plasma Spectrochemistry CY - Fort Myers, FL, USA DA - 2010-01-04 PY - 2010 AN - OPUS4-20838 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Roos, P.H. A1 - Jakubowski, Norbert T1 - Methods for the discovery of low-abundance biomarkers for urinary bladder cancer in biological fluids N2 - For the study of bladder cancer and the identification of respective tumor markers, blood and, in particular, urine constitute suitable sources of biological material, while both harboring their specific challenges for analytics concerning low-abundance biomarkers. Dissolved proteins and nucleic acids as well as cells and cell-bound molecules can be the analytes. In urine, exfoliated bladder tumor cells have to be identified and in blood, circulating tumor cells have to be detected among huge amounts of other cells. For the detection of both low-abundance cells and molecules, their specific enrichment prior to analysis is advantageous or even necessary. Adapted methods for the analysis of proteomes and subproteomes by 2D-gel electrophoresis, multidimensional chromatography and antibody arrays are discussed. Analysis of nucleic acid-based markers exploits the high amplification power of PCR and modified PCR combined with previous (subtransciptomes) or subsequent (microarray) enrichment to sensitively and specifically detect markers. DNA mutations, DNA-methylation status and apoptotic DNA fragments, as well as levels of ribonucleic acids including microRNAs, can be analyzed by means of these methods. Finally, the challenge of identifying circulating tumor cells and assigning them to their original tissue is critically discussed. PY - 2010 DO - https://doi.org/10.4155/BIO.09.174 SN - 1757-6180 VL - 2 IS - 2 SP - 295 EP - 309 PB - Future Science Group CY - London AN - OPUS4-20981 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Nießner, R. A1 - Broekaert, J.A. A1 - Bron, M. A1 - Einax, J. A1 - Emons, H. A1 - Haisch, C. A1 - Huber, C. A1 - Jakubowski, Norbert A1 - Knopp, D. A1 - Popp, J. A1 - Schäferling, M. T1 - Analytische Chemie 2008/2009 PY - 2010 DO - https://doi.org/10.1002/nadc.201063123 SN - 1439-9598 SN - 1521-3854 VL - 58 IS - 3 SP - 223 EP - 235 PB - Wiley-VCH Verl. CY - Weinheim AN - OPUS4-21177 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Heritage of Prof. Dr. Ing. Klaus Gustav Heumann: Selected Highlights T2 - 2010 Winter Conference CY - Fort Myers, FL, USA DA - 2010-01-04 PY - 2010 AN - OPUS4-21161 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - GEN A1 - Jakubowski, Norbert A1 - Prohaska, T. A1 - Roos, P.H. ED - Beauchemin, D. ED - Matthews, D. T1 - Polyatomic ions with double-focusing magnet sector mass spectrometers KW - ICP-MS KW - Sektorfeldgeräte PY - 2010 SN - 978-0-08-043804-7 VL - 5 SP - 132 EP - 150 PB - Elsevier CY - Oxford, UK AN - OPUS4-22237 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Roos, P.H. A1 - Jakubowski, Norbert T1 - Cytochromes - fascinating molecular machines N2 - A huge number of functions and reactions can be performed by proteins merely based on their amino acids and their spatial arrangement which results in a specific geometric and electronic environment. The diversity and efficiency of functions, however, can be increased by protein-bound cofactors which fulfil their task in close interaction with the surrounding amino acid side chains. Among these cofactors, metals play an important role. They can be bound to proteins directly via specific amino acids such as cysteine and histidine, for example in zinc finger proteins or in iron sulfur cluster proteins. Another possibility is their integration into the 3D-structure of a protein via a prosthetic group. Porphyrine or porphyrine-like molecules with copper or iron as central bound metal ions constitute important structures in this respect. PY - 2011 DO - https://doi.org/10.1039/c1mt90012a SN - 1756-5901 SN - 1756-591X VL - 3 SP - 316 EP - 318 PB - RSC Publ. CY - Cambridge AN - OPUS4-23830 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Neue Anwendungen der anorganischen Massenspektrometrie in den Lebens- und Materialwissenschaften T2 - Eingeladener Vortrag, Fakultätskolloquium, Technische Universität Bergakademie Freiberg CY - Freiberg, Germany DA - 2011-02-15 PY - 2011 AN - OPUS4-23809 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert A1 - Tempez, A. A1 - Chapon, P. A1 - Bordel, N. A1 - Pisonero, J. A1 - Lobo, L. T1 - A new Glow Discharge Time of Fligh Mass Spectrometer for Bulk Analysis and Depth Profiling T2 - PITTCON 2010, eingeladener Vortrag CY - Atlanta, GA, USA DA - 2011-03-13 PY - 2011 AN - OPUS4-23810 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Moreno-Gordaliza, E. A1 - Esteban-Fernández, D. A1 - Giesen, Charlotte A1 - Lehmann, K. A1 - Lázaro, A. A1 - Tejedor, A. A1 - Scheler, C. A1 - Canas, B. A1 - Jakubowski, Norbert A1 - Linscheid, M.W. A1 - Gómez-Gómez, M.M. T1 - LA-ICP-MS and nHPLC-ESI-LTQ-FT-MS/MS for the analysis of cisplatin-protein complexes separated by two dimensional gle elctrophoresis in biological samples N2 - A method for the analysis of Pt–protein complexes in biological samples, previously subjected to cisplatin treatment, has been developed. Proteins were separated by gel electrophoresis, and those bound to Pt were detected with high sensitivity by LA-ICP-(SF)-MS. Pt-containing spots were in-gel digested with trypsin, and the peptides produced identified using nHPLC-ESI-LTQ-FT-MS/MS. The influence of protein separation conditions, staining and gel processing prior to laser ablation on Pt–protein bonds preservation have been evaluated using standard proteins incubated with cisplatin. 2-DE separation under non-reducing conditions followed by either Coomassie blue brilliant or silver staining is appropriate for Pt–protein complexes, achieving a good separating resolution of the proteins in biological samples. Direct LA-ICP-MS analysis of glycerol-treated dried gels for Pt–protein monitoring resulted in better sensitivity, more reliable relative Pt signals and a simpler and less time-consuming approach compared to the analysis of blotted membranes. Ablation of gels allowed tackling protein identification of Pt-spots in the remaining non-ablated material in the gel, making it unnecessary to run several gels in parallel for separate Pt detection and protein identification. By using this approach, Pt coordinated to proteins, such as α-2-macroglobulin, transferrin, albumin or hemoglobin, was detected in the serum from a rat treated in vivo with cisplatin after nrSDS-PAGE separation. Furthermore, the first complete LA-ICP-MS metalloprotein contour map in a 2-DE gel has been produced, in this case for the detection of Pt–protein complexes in renal proximal tubule epithelial cells (RPTECs) incubated with cisplatin. Several proteins were identified in those spots containing Pt, which may have a connection with the drug-induced nephrotoxicity mainly affecting this cell type in the kidney. PY - 2012 DO - https://doi.org/10.1039/c2ja30016h SN - 0267-9477 SN - 1364-5544 VL - 27 IS - 9 SP - 1474 EP - 1483 PB - Royal Society of Chemistry CY - London AN - OPUS4-26562 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -