{"id":690,"date":"2022-05-05T12:32:49","date_gmt":"2022-05-05T10:32:49","guid":{"rendered":"https:\/\/biochemie-nat.cms.rrze.uni-erlangen.de\/?page_id=690"},"modified":"2024-04-15T13:16:02","modified_gmt":"2024-04-15T11:16:02","slug":"ag-joerg-hofmann","status":"publish","type":"page","link":"https:\/\/www.biochemie.nat.fau.de\/en\/ag-joerg-hofmann\/","title":{"rendered":"Bioanalytics &#8211; Group J\u00f6rg Hofmann"},"content":{"rendered":"<p>The <strong>Bioanalytics group<\/strong> established a <strong>Metabolomics<\/strong> technology platform and a <strong>Proteomics<\/strong> platform enabling the qualitative and quantitative analysis of biomolecules. This includes targeted and untargeted metabolite profiling. The Metabolomics branch consists of the following equipment:<\/p>\n<h2>Metabolomics platform<\/h2>\n<h3>LC\/MS-MS System<\/h3>\n<ul>\n<li>ABI\/MDS-Sciex QTRAP-3200 (Mass Spectrometer)<\/li>\n<li>Dionex ICS-3000 Ion Exchange HPLC (metabolite analysis)<\/li>\n<li>Dionex ULTIMATE-3000 HPLC (metabolite analysis)<\/li>\n<\/ul>\n<h4>Other instruments<\/h4>\n<ul>\n<li>Dionex SUMMIT P680 HPLC (metabolite analysis)<\/li>\n<li>Shimadzu GCMS-QP2010S mit GC-2010 (Fatty acid analysis)<\/li>\n<li>\u00c4kta Purifier HPLC (Protein purification)<\/li>\n<li>Pharmacia P-800 FPLC (low pressure LC)<\/li>\n<li>Packard Flow Scintillation Analyser (Radioactive molecules)<\/li>\n<li>BIO-TEK ELISA Multiwell microplate photometer (Vis fluorescence)<\/li>\n<li>Goebel UVIKON-XL Spectrophotometer (UV\/Vis)<\/li>\n<\/ul>\n<h4>Access<\/h4>\n<p>Prior to use the analysis platform the requirements of our regulations \u00a0 <a href=\"https:\/\/biochemie-nat.cms.rrze.uni-erlangen.de\/files\/2022\/05\/nutzunglsbiochemie2016metabolomics.pdf\"><strong>(PDF, german)<\/strong><\/a>\u00a0 apply and a sample submission form which is available on request must be filled in and sent via e-mail to: \u00a0 <em>joerg.hofmann (at) fau.de<\/em><\/p>\n<p>The core of our <strong>LC-MS\/MS<\/strong> system represents the <em>QTrap-3200<\/em> which is a versatile hybrid mass spectrometer. It combines the features of three cascaded mass filters (Triple-Quadrupole) needed for precise quantification with the concept of a linear iontrap enabling to derive information about quality (structure determination). This combination cares for an enhanced sensitivity. The Triple-Quadrupole together with other technical features (eg. N2-curtain) allows the analysis of relatively crude samples. For sensitive molecules we can adjust the harshness of ionisation by switching between different ion-sources ( Gas assisted Electro Spray Ionisation (<strong>ESI<\/strong>), Atmospheric pressure chemical ionisation (<strong>APCI<\/strong>) ). The ion source receives the stream of analytes after separation by the <em>ICS-3000<\/em>&#8211; ion chromatography- or the <em>Ultimate-3000<\/em> HPLC system.<br \/>\nThe inert <em>ICS-3000<\/em> IC is exceptionally suited for <strong>ion exchange chromatography<\/strong> e.g. of <strong>carbohydrates<\/strong> or <strong>phosphorylated intermediates<\/strong>. It generates a &#8220;reagent-free&#8221; eluent required for mass spectrometry and has additional detectors (conductivity- and 3D amperometry detector) e.g. for the screening of <strong>sugars<\/strong>. In combination with the <em>QTrap<\/em> mass spectrometer we can use this device for targeted metabolite profiling with higher dynamic range and a broad spectrum of compounds.<br \/>\nCombining the <em>Ultimate 3000<\/em> HPLC with an <em>UV\/Vis Photodiode Array Detector (PDA)<\/em> results in a sophisticated 3D-visualization of fluorescence signals allowing easy identification and sensitve quantification of e.g. <strong>pigments<\/strong>, such as <strong>carotenoids<\/strong> or <strong>chlorophyll<\/strong>.<br \/>\nWe have an additional HPLC system (<em>Dionex SUMMIT<\/em>) with a fluorescence detector suitable for the sensitive measurement of compounds like <strong>tocopherol<\/strong> and (derivatized) <strong>amino acids<\/strong> after Reversed Phase (RP) Chromatography.<br \/>\nWe are also currently improving methods for GC\/MS-based determination of <strong>fatty acids<\/strong>.<br \/>\nOur newly installed laboratory is further equipped with various supporting devices. Two types of absorption photometers are used to measure e.g. <strong>sugar<\/strong> concentration: 96-well ELISA Plate Readers with washer and fluorescence-detector facilitate quick simultaneous analyses of multiple samples. UV\/Vis Spectrophotometers allow more sensitive absorption measurements.<br \/>\nIn combination with our Flow Scintillation Analyser after chromatographic separation, we can detect <strong>radioactively labeled compounds<\/strong>. Furthermore we have installed a cold lab with an <em>\u00c4kta purifier<\/em> and a low pressure FPLC sytem for the separation of <strong>proteins<\/strong>.<br \/>\nThis equipment allows the parallel investigation of metabolites in complex mixtures and supports projects aimed at investigating plant-microbe interactions and primary plant metabolism ( <a href=\"https:\/\/biochemie-nat.cms.rrze.uni-erlangen.de\/ag-sophia-sonnewald\/\">AG S.Sonnewald<\/a>, <a href=\"https:\/\/biochemie-nat.cms.rrze.uni-erlangen.de\/ag-uwe-sonnewald\/\">AG U.Sonnewald<\/a> ). Furthermore, plant-cell-to-cell connections (plasmodesmata) are analysed at the biochemical and molecular level.<\/p>\n<h4>Proteomics platform<\/h4>\n<div class=\"feedcontent\">\n<p>As part of <strong>Zentralprojekt Z1<\/strong> in the <strong>Sonderforschungsbereich SFB796<\/strong> a Proteomics lab was established in 2014. Identification and analysis of proteins and protein complexes with their post-translational modifications can be performed with mass spectrometry instruments.<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\" wp-image-490 alignright\" src=\"https:\/\/biochemie-nat.cms.rrze.uni-erlangen.de\/files\/2022\/04\/protlogo1_200_69.png\" alt=\"\" \/><\/p>\n<p>This platform consists of <strong>Ultimate 3000 nano-HPLC<\/strong> and <strong>Orbitrap Fusion Tribrid<\/strong> systems designed for proteomic bioanalytics research. Optimized procedures of sample preparation and analysis enable us to work effectively with purified fractions, intact proteins, SDS-PAGE bands\/spots, immunoprecipitated proteins (on\/off-beads) for the identification of<\/p>\n<ul>\n<li>\u00a0 Interacting partners<\/li>\n<li>\u00a0 PTMs<\/li>\n<li>\u00a0 Differences at protein level (e.g. wild type vs. mutation, transfection, differential expression, stress influence)<\/li>\n<li>\u00a0 Proteome\/Secretome studies<\/li>\n<\/ul>\n<p>Special projects like <strong>Biomarker discovery<\/strong>, <strong>quantification<\/strong> (labelled\/label-free) and <strong>phosphoproteome analysis<\/strong> are under development.<\/p>\n<p>Operation and management of the platform is conducted by \u00a0 <strong>J\u00f6rg Hofmann<\/strong><\/p>\n<p>For further details please contact: \u00a0 <strong> joerg.hofmann (at) fau.de <\/strong> , \u00a0 <strong> uwe.sonnewald (at) fau.de <\/strong><\/p>\n<p><strong><em>Sample submission form<\/em> \u00a0 to apply for a measurement:<\/strong><br \/>\nDownload this <a href=\"https:\/\/biochemie-nat.cms.rrze.uni-erlangen.de\/files\/2022\/05\/samplesubmissionform_27-05-2015_1.pdf\"><strong>PDF<\/strong><\/a> into a file, fill it in, print it into a PDF file again and send it by e-mail.<\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>The Bioanalytics group established a Metabolomics technology platform and a Proteomics platform enabling the qualitative and quantitative analysis of biomolecules. This includes targeted and untargeted metabolite profiling. The Metabolomics branch consists of the following equipment: Metabolomics platform LC\/MS-MS System ABI\/MDS-Sciex QTRAP-3200 (Mass Spectrometer) Dionex ICS-3000 Ion Exchange HPLC (metabolite analysis) Dionex ULTIMATE-3000 HPLC (metabolite analysis) [&hellip;]<\/p>\n","protected":false},"author":859,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_rrze_cache":"enabled","_access_permission":"","_rrze_multilang_single_locale":"en_US","_rrze_multilang_single_source":"https:\/\/biochemie-nat.cms.rrze.uni-erlangen.de\/?page_id=123","footnotes":""},"page_category":[22],"page_tag":[],"workflow_usergroup":[],"class_list":["post-690","page","type-page","status-publish","hentry","page_category-ags","en-US"],"_links":{"self":[{"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/pages\/690","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/users\/859"}],"replies":[{"embeddable":true,"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/comments?post=690"}],"version-history":[{"count":10,"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/pages\/690\/revisions"}],"predecessor-version":[{"id":1569,"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/pages\/690\/revisions\/1569"}],"wp:attachment":[{"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/media?parent=690"}],"wp:term":[{"taxonomy":"page_category","embeddable":true,"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/page_category?post=690"},{"taxonomy":"page_tag","embeddable":true,"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/page_tag?post=690"},{"taxonomy":"workflow_usergroup","embeddable":true,"href":"https:\/\/www.biochemie.nat.fau.de\/wp-json\/wp\/v2\/workflow_usergroup?post=690"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}