{"id":1069,"date":"2024-12-09T09:19:52","date_gmt":"2024-12-09T09:19:52","guid":{"rendered":"https:\/\/hplc2025-bruges.org\/?page_id=1069"},"modified":"2025-04-24T07:16:10","modified_gmt":"2025-04-24T07:16:10","slug":"tutorials","status":"publish","type":"page","link":"https:\/\/hplc2025-bruges.org\/?page_id=1069","title":{"rendered":"Tutorials"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-page\" data-elementor-id=\"1069\" class=\"elementor elementor-1069\" data-elementor-post-type=\"page\">\n\t\t\t\t<div class=\"elementor-element elementor-element-8b832fa e-flex e-con-boxed e-con e-parent\" data-id=\"8b832fa\" data-element_type=\"container\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t<div class=\"elementor-element elementor-element-e3a7476 elementor-widget elementor-widget-heading\" data-id=\"e3a7476\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">Tutorials<\/h2>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-8c6728e e-flex e-con-boxed e-con e-parent\" data-id=\"8c6728e\" data-element_type=\"container\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t<div class=\"elementor-element elementor-element-c23d1c1 elementor-widget elementor-widget-spacer\" data-id=\"c23d1c1\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-51b630d e-con-full e-flex e-con e-parent\" data-id=\"51b630d\" data-element_type=\"container\">\n\t\t<div class=\"elementor-element elementor-element-c579c31 e-grid e-con-full e-con e-child\" data-id=\"c579c31\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-ef10b4f elementor-widget elementor-widget-image\" data-id=\"ef10b4f\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" src=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/elementor\/thumbs\/Lars-Nilsson-2-qzy36548vk68d7ud89yazzfjviro1v6ec90v72h5ns.jpg\" title=\"Lars Nilsson 2\" alt=\"Lars Nilsson 2\" loading=\"lazy\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-a1edbb1 elementor-widget elementor-widget-text-editor\" data-id=\"a1edbb1\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00a0 \u00a0<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-212566d e-con-full e-flex e-con e-child\" data-id=\"212566d\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-4277119 elementor-widget elementor-widget-text-editor\" data-id=\"4277119\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>TUTORIAL 01 &#8211; Wed June 18 &#8211;\u00a0 08h30 \/ 09h15<\/p><p>LARS <strong>NILSSON<\/strong><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-62a58fc elementor-widget elementor-widget-heading\" data-id=\"62a58fc\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">Asymmetric flow field-flow fractionation (AF4) coupled to synchrotron small angle x-ray scattering (SAXS) \u2013 A new tool for characterization of proteins and nanoparticles<\/h2>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-9ca88c7 e-con-full e-flex e-con e-child\" data-id=\"9ca88c7\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-8dad92b elementor-widget elementor-widget-tp-unfold\" data-id=\"8dad92b\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e056c92 tp-ca-center \" data-id=\"plus6a9679e056c92\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Speaker Biosketch\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Lars Nilsson<\/strong> has a background in surface and colloid science, receiving his PhD from Lund University, Sweden in 2007. The thesis subject was emulsification and macromolecular emulsifiers. The work brought him into contact with field-flow fractionation under the mentorship of Professor emeritus Karl-Gustav Wahlund, who pioneered asymmetric flow field flow fractionation (AF4). After a period in the biotech industry, he returned to Lund University and increasingly engaged in AF4 methodology and applications. In 2012 a spinoff company offering AF4 and complementary techniques as a contract service was founded. In 2020 he became full Professor in Formulation Technology at Lund University. His current research focus is on the formulation of therapeutic proteins, lipid nanoparticles and biomolecular corona formation on nanoparticles as well as development and application of AF4 methodology. This involves the combination of AF4 with synchrotron small angle x-ray scattering (SAXS) and small angle neutron scattering (SANS).<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Speaker Biosketch <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-33c19df elementor-widget elementor-widget-tp-unfold\" data-id=\"33c19df\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e057374 tp-ca-center \" data-id=\"plus6a9679e057374\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Tutorial Description\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Asymmetric Flow-Field Flow Fractionation (AF4) for the characterization of biopharmaceuticals<\/strong><\/p><p>Asymmetric flow field-flow fractionation (AF4) is a powerful size-based separation technique that has been used for several decades in the analysis and characterization of biopharmaceuticals. Unlike traditional chromatographic methods, AF4 operates without a stationary phase, making it particularly advantageous for analyzing delicate or complex macromolecules and nanoparticles that may otherwise be affected by interactions with a solid matrix. AF4 has proven especially useful when more widely used separation techniques, such as size-exclusion chromatography (SEC), encounter limitations. SEC, while widely employed, may struggle with samples that include broad or multimodal size distributions, particularly those containing large analytes or aggregates. In contrast, AF4 provides a broader dynamic range, allowing for the effective separation of nanoparticles, protein aggregates, and other macromolecular complexes that exceed the size limits of SEC. This makes AF4 an invaluable tool for researchers working with biopharmaceutical formulations, where precise size characterization is critical for ensuring product efficacy and safety.<\/p><p>Early applications of AF4 in biopharmaceuticals primarily focused on the separation and analysis of protein oligomers and aggregates, which are key factors in drug stability, efficacy and immunogenicity. In recent years, there has been a significant surge of interest in AF4 across a broader scientific community, largely driven by the growing emphasis on nanoparticle-based drug delivery systems. Nanoparticles, including lipid and polymeric nanoparticles, and viral vectors, play a crucial role in modern drug development. The ability of AF4 to accurately fractionate nanoparticle size distributions is positioning it as a key analytical tool in this rapidly evolving field.<\/p><p><strong>What to Expect from This Tutorial<\/strong><\/p><p>This tutorial will provide an overview of the fundamental principles of AF4, explaining how the technique operates and highlighting its key advantages and limitations compared to other separation methods. Additionally, example applications of AF4 within biopharmaceutical research will be explored, showcasing its versatility.<\/p><p>By the end of this tutorial, attendees will have gained an insight into AF4\u2019s role in biopharmaceutical analysis and how it can be effectively applied to address challenges that other separation techniques may struggle to overcome.<\/p><p>- - -<\/p><p><strong>Lars Nilsson<\/strong> -\u00a0Dept of Process and Life Science Engineering, Lund University, Lund, Sweden\u00a0 <a href=\"mailto:lars.nilsson@ple.lth.se\">lars.nilsson@ple.lth.se<\/a><\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Tutorial Description <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-bd90a4d e-flex e-con-boxed e-con e-parent\" data-id=\"bd90a4d\" data-element_type=\"container\" data-settings=\"{&quot;background_background&quot;:&quot;classic&quot;}\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t<div class=\"elementor-element elementor-element-43366d2 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"43366d2\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-f4975c7 e-con-full e-flex e-con e-parent\" data-id=\"f4975c7\" data-element_type=\"container\">\n\t\t<div class=\"elementor-element elementor-element-3fdcb91 e-grid e-con-full e-con e-child\" data-id=\"3fdcb91\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-368431f elementor-widget elementor-widget-image\" data-id=\"368431f\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" src=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/elementor\/thumbs\/Tim-Causon-2-scaled-qzy36548vk68d7ud89yazzfjviro1v6ec90v72h5ns.jpg\" title=\"Tim Causon 2\" alt=\"Tim Causon 2\" loading=\"lazy\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-d17c9fb elementor-widget elementor-widget-text-editor\" data-id=\"d17c9fb\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00a0 \u00a0<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-c419cc4 e-con-full e-flex e-con e-child\" data-id=\"c419cc4\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-f7abbfc elementor-widget elementor-widget-text-editor\" data-id=\"f7abbfc\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>TUTORIAL 02 &#8211; Wed June 18 &#8211; 10h45 \/ 11h30<\/p><p>TIM <strong>CAUSON<\/strong><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-9661334 elementor-widget elementor-widget-heading\" data-id=\"9661334\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">Ion mobility-mass spectrometry and its hyphenation<\/h2>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-63e1360 e-con-full e-flex e-con e-child\" data-id=\"63e1360\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-0b5c6ff elementor-widget elementor-widget-tp-unfold\" data-id=\"0b5c6ff\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e05a0cf tp-ca-center \" data-id=\"plus6a9679e05a0cf\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Speaker Biosketch\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Tim Causon<\/strong> is an Associate Professor at Institute of Analytical Chemistry, BOKU University in Vienna, Austria. He is the co-ordinator of the MSCA Doctoral Network \u201c<em>MobiliTraIN<\/em>\u201d (<em>Ion Mobility Mass Spectrometry Training Network, www.mobilitrain.eu<\/em>). His main areas of research are liquid chromatography, molecular mass spectrometry, and ion mobility-mass spectrometry (IM-MS) for diverse analytical method development challenges and fundamental studies of ionisation behaviour of small molecular ions. Current research topics include development of methods for optimizing upstream (e.g., microbial cell factories) and downstream (e.g., purification) ends of the bioprocessing continuum, high-resolution ion mobility separation using Structures for Lossless ion Manipulation (SLIM), and coordinated efforts toward harmonization of IM-MS measurement and reporting standards.<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Speaker Biosketch <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-e46c131 elementor-widget elementor-widget-tp-unfold\" data-id=\"e46c131\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e05a58f tp-ca-center \" data-id=\"plus6a9679e05a58f\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Tutorial Description\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Introduction to IM theory, hyphenated IM-MS instrument technology, analytical applications<\/strong><\/p><p>As a separation technique, ion mobility (IM) complements existing analytical methods as it involves millisecond-timescale separations of gas phase ions according to their structure and not only their mass\/charge. Combined with mass spectrometry (MS), IM-MS is now a next generation technology that can be applied to very diverse analytical applications especially when hyphenated with liquid-phase separation techniques. This tutorial will introduce the basics of IM theory, the differences between LC-MS and LC-IM-MS acquisition and show how collision cross sections (CCS) from IM-MS measurements can be used to support the identification of unknowns. Principles of the most important IM-MS technologies will be introduced, and hyphenated application examples from diverse fields (e.g., food, metabolomics, environmental analysis) will be used to highlight how IM-MS can be successfully applied in analytical methods.<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Tutorial Description <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-4c79788 e-con-full e-flex e-con e-parent\" data-id=\"4c79788\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-dbedd03 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"dbedd03\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-cf9d855 e-grid e-con-full e-con e-child\" data-id=\"cf9d855\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-0291410 elementor-widget elementor-widget-image\" data-id=\"0291410\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" src=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/elementor\/thumbs\/Szabolcs-Fekete-qzy36548vk68d7ud89yazzfjviro1v6ec90v72h5ns.jpeg\" title=\"Szabolcs Fekete\" alt=\"Szabolcs Fekete\" loading=\"lazy\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-7a3dd39 elementor-widget elementor-widget-image\" data-id=\"7a3dd39\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" width=\"150\" height=\"150\" src=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/2024\/12\/93E6BB67-C5CA-47C2-8343-943E7BDA9BDB_1_201_a-150x150.jpeg\" class=\"attachment-thumbnail size-thumbnail wp-image-1095\" alt=\"\" srcset=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/2024\/12\/93E6BB67-C5CA-47C2-8343-943E7BDA9BDB_1_201_a-150x150.jpeg 150w, https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/2024\/12\/93E6BB67-C5CA-47C2-8343-943E7BDA9BDB_1_201_a-300x300.jpeg 300w, https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/2024\/12\/93E6BB67-C5CA-47C2-8343-943E7BDA9BDB_1_201_a-768x768.jpeg 768w, https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/2024\/12\/93E6BB67-C5CA-47C2-8343-943E7BDA9BDB_1_201_a-700x700.jpeg 700w, https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/2024\/12\/93E6BB67-C5CA-47C2-8343-943E7BDA9BDB_1_201_a.jpeg 774w\" sizes=\"(max-width: 150px) 100vw, 150px\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-ccdf4ef e-con-full e-flex e-con e-child\" data-id=\"ccdf4ef\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-3c8c508 elementor-widget elementor-widget-text-editor\" data-id=\"3c8c508\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>TUTORIAL 03 \u2013 Wed June 18 \u2013 14h15 \/ 15h00<\/p><p>SZABOLCS\u00a0<strong>FEKETE<\/strong><br \/>MATEUSZ\u00a0<strong>IMIOLEK<\/strong><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-dc4f8c5 elementor-widget elementor-widget-heading\" data-id=\"dc4f8c5\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">Chromatographic strategies for the analytical characterization of cell and gene therapy products<\/h2>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-3bd187e e-con-full e-flex e-con e-child\" data-id=\"3bd187e\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-f8094be elementor-widget elementor-widget-tp-unfold\" data-id=\"f8094be\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e05ce33 tp-ca-center \" data-id=\"plus6a9679e05ce33\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Speaker Biosketch\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Szabolcs Fekete<\/strong>, Consulting Scientist, Cell and Gene Therapy, Consumables at Waters Corporation<\/p><p>Szabolcs Fekete received his PhD in Analytical Chemistry from the Technical University of Budapest. He worked in analytical R&amp;D in the pharmaceutical industry for 10 years, then joined the University of Geneva in Switzerland as a research associate. He joined Waters Corporation in 2021 and is now a consulting scientist. His current interests include separations of new chemical modalities, fundamentals of chromatography, column technology and new method development approaches. He has co-authored ~200 publications (including journal articles, book chapters and handbooks).<\/p><p><strong>Mateusz<\/strong>\u00a0joined Waters in 2022 as an Application Scientist where he has been studying challenges associated with characterizing gene therapeutics and ensuring that new Waters products are fit for this purpose. Before that, he completed his academic training (PhD + postdoc) in bioorganic chemistry, which allows him to apply detailed molecular insights during development of new columns and assays carefully designed for such analytes.<\/p><p>Working in a close collaboration with the Institute of Pharmaceutical Sciences of Western Switzerland, he has been advancing LC-based methods (SEC, IP-RP, IEX) for analysis of new modality biopharmaceuticals with a recent focus on AAV, mRNA and LNP chromatographic separations. He is motivated to accelerate market introduction of safe and efficacious cell and gene therapy drugs via providing better analytical tools.<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Speaker Biosketch <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-1ea9b89 elementor-widget elementor-widget-tp-unfold\" data-id=\"1ea9b89\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e05d2ed tp-ca-center \" data-id=\"plus6a9679e05d2ed\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Tutorial Description\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Chromatographic strategies for the analytical characterization of genetic medicine drug substances<\/strong><\/p><p>Szabolcs Fekete, Mateusz Imiolek<\/p><p>Waters Corporation, Geneva, Switzerland<\/p><p>The rapid evolution of oligonucleotide (ON) and nucleic acid therapeutics has introduced complex analytical challenges, particularly in impurity profiling and structural characterization. Issues such as 5\u2032-methylcytidine deamination, residual single-strand content in duplex formulations, the increased complexity of conjugated ONs, and the formation of aggregates necessitate more advanced chromatographic solutions.<\/p><p>This tutorial presents a suite of cutting-edge liquid chromatography (LC) strategies developed to address these challenges, with a focus on improving both resolution and throughput. We showcase innovations in ion-pair reversed-phase liquid chromatography (IP-RPLC), including segmented (concave\/logarithmic) gradients to optimize peak distribution, enhanced selectivity through dynamic ion-pairing transitions, and the use of ultra-short columns to minimize analysis time and solvent consumption.<\/p><p>Hydrophilic interaction chromatography (HILIC) is also explored as a robust, ion-pair-free alternative, with systematically optimized conditions for the separation of ON sequence and length variants. Key parameters such as ionic strength, temperature, gradient profile, and injection technique will be discussed.<\/p><p>Additionally, we examine diverse elution strategies for anion exchange chromatography (AEX), including salt gradients, ion-pair gradients (IPAX), combined ion-pair\/salt gradients, and pH gradients. The potential of weak AEX phases for analyzing phosphorothioate antisense oligonucleotide (ASO) impurities, such as deaminated species, will be demonstrated, along with \u201csalt plug injection\u201d techniques to improve separation performance.<\/p><p>We further highlight advances in ultra-wide pore size exclusion chromatography (SEC), where sub-3 \u00b5m particle columns with &gt;500 \u00c5 pore sizes provide high-resolution separations of large nucleic acid modalities \u2014 including mRNA and plasmids \u2014 with enhanced compatibility for multiangle light scattering (MALS) detection. In parallel, slalom chromatography is emerging as a powerful method for the separation and mass analysis of long double-stranded DNA, offering a modern, LC-based alternative to traditional gel electrophoresis.<\/p><p>These approaches deliver improved sensitivity, quantification and accelerating the development of advanced gene and cell therapies.<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Tutorial Description <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-8b85bf6 e-flex e-con-boxed e-con e-parent\" data-id=\"8b85bf6\" data-element_type=\"container\" data-settings=\"{&quot;background_background&quot;:&quot;classic&quot;}\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t<div class=\"elementor-element elementor-element-9ba8c21 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"9ba8c21\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-2dfe145 e-con-full e-flex e-con e-parent\" data-id=\"2dfe145\" data-element_type=\"container\">\n\t\t<div class=\"elementor-element elementor-element-cb9913d e-grid e-con-full e-con e-child\" data-id=\"cb9913d\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-27e4298 elementor-widget elementor-widget-image\" data-id=\"27e4298\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" src=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/elementor\/thumbs\/Elia-Psillakis-qzy36548vk68d7ud89yazzfjviro1v6ec90v72h5ns.jpeg\" title=\"Elia Psillakis\" alt=\"Elia Psillakis\" loading=\"lazy\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-8bfd3ec elementor-widget elementor-widget-text-editor\" data-id=\"8bfd3ec\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00a0 \u00a0<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-6826d18 e-con-full e-flex e-con e-child\" data-id=\"6826d18\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-92490ae elementor-widget elementor-widget-text-editor\" data-id=\"92490ae\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>TUTORIAL 4 &#8211; Wed June 18 &#8211; 16h30 \/ 17h15<\/p><p>ELIA <strong>PSILAKIS<\/strong><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-37e3dd9 elementor-widget elementor-widget-heading\" data-id=\"37e3dd9\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">How to quantify the greenness of your methods?<\/h2>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-64e8398 e-con-full e-flex e-con e-child\" data-id=\"64e8398\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-412bec2 elementor-widget elementor-widget-tp-unfold\" data-id=\"412bec2\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e05fbee tp-ca-center \" data-id=\"plus6a9679e05fbee\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Speaker Biosketch\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Elefteria (Elia) Psillakis<\/strong> is a Water Chemistry Professor at the School of Chemical and Environmental Engineering, Technical University of Crete, Greece. In 2007, she received a Fulbright Award as a visiting Professor at CALTECH, USA. She was featured in the Top 100 Power List 2021 of the magazine \u201cThe Analytical Scientist\u201d and has received three \u201cTop cited article\u201d awards. She has published more than 100 papers that have received ~10000 citations. She is Editor-in-Chief of \u201cAdvances in Sample Preparation\u201d (Elsevier) and the Head of the Sample Preparation Network of the EuChemS-DAC. She is also the Founder and Director of ExtraTECH Analytical Solutions, a Spin-Off Company of the Technical University of Crete. From 2014-2016, she was the Vice Rector of Academic Affairs and Research at the Technical University of Crete<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Speaker Biosketch <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-603d728 elementor-widget elementor-widget-tp-unfold\" data-id=\"603d728\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e060049 tp-ca-center \" data-id=\"plus6a9679e060049\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Tutorial Description\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p>This tutorial introduces participants to AGREE and AGREEprep, two user-friendly green metrics designed to quantify the greenness of analytical and sample preparation methods. Attendees will explore the concepts behind these tools, learn how to apply them, gain hands-on experience, interpret their outputs, and use the results to improve the green profile of their methods. Whether developing new protocols or optimizing existing ones, this session offers practical guidance for embedding greenness into the analytical workflow, gaining valuable insight into how individual criteria contribute to the final evaluation from the assessor\u2019s perspective, and assisting in identifying specific strengths and weaknesses of analytical methods.<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Tutorial Description <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-48bc57b e-flex e-con-boxed e-con e-parent\" data-id=\"48bc57b\" data-element_type=\"container\" data-settings=\"{&quot;background_background&quot;:&quot;classic&quot;}\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t<div class=\"elementor-element elementor-element-289a000 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"289a000\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-fdabdf2 e-con-full e-flex e-con e-parent\" data-id=\"fdabdf2\" data-element_type=\"container\">\n\t\t<div class=\"elementor-element elementor-element-6ac391b e-grid e-con-full e-con e-child\" data-id=\"6ac391b\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-26fafb6 elementor-widget elementor-widget-image\" data-id=\"26fafb6\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" src=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/elementor\/thumbs\/a7d3d111816794c072800624b40be395cd120f5c-245x245-1-qzy36548vk68d7ud89yazzfjviro1v6ec90v72h5ns.webp\" title=\"a7d3d111816794c072800624b40be395cd120f5c-245\u00d7245\" alt=\"a7d3d111816794c072800624b40be395cd120f5c-245x245\" loading=\"lazy\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-cb79830 elementor-widget elementor-widget-text-editor\" data-id=\"cb79830\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00a0 \u00a0<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-85f9a8d e-con-full e-flex e-con e-child\" data-id=\"85f9a8d\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-b96f3fe elementor-widget elementor-widget-text-editor\" data-id=\"b96f3fe\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>TUTORIAL 05 &#8211; Thu June 19 &#8211; 08h30 \/ 09h15<\/p><p>PETER <strong>SCHOENMAKERS<\/strong><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-a6952d3 elementor-widget elementor-widget-heading\" data-id=\"a6952d3\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">No molecule is above the law (no matter how big and strong they are)<\/h2>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-3a2fac4 e-con-full e-flex e-con e-child\" data-id=\"3a2fac4\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-d35fe49 elementor-widget elementor-widget-tp-unfold\" data-id=\"d35fe49\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e0629c6 tp-ca-center \" data-id=\"plus6a9679e0629c6\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Speaker Biosketch\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Peter Schoenmakers<\/strong> has been a full-time professor in Analytical Chemistry at the University of Amsterdam since 2002 \u2013 with emeritus status since 2021. His has worked on many aspects of one- and two-dimensional chromatography, with a special interest in the characterization of macromolecules. He has seen method-optimization and artificial-intelligence tools emerge some forty years ago, with only gradual progress until recently. Finally, we are starting to see exciting progress in method development for LC.<\/p><p>Together with Bob Pirok, Peter Schoenmakers has recently completed a new textbook on Analytical Separation Science, which is to be launched at HPLC2025 Bruges. He is also the Education Director of COAST, The Netherlands\u2019 public-private-partnership organization on analytical chemistry. \u00a0From 2005 to 2020 he was an editor of the Journal of Chromatography A. He has received a number of international awards, most recently the ACS Award for Chromatography (2022) and the LCGC lifetime-achievement award (2023).<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Speaker Biosketch <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-e26d0c1 elementor-widget elementor-widget-tp-unfold\" data-id=\"e26d0c1\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e062e2c tp-ca-center \" data-id=\"plus6a9679e062e2c\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Tutorial Description\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>No molecule is above the law (no matter how big and strong they are)<\/strong><\/p><p>The same fundamental chromatographic laws apply to all molecules - small (i.e. low molecular weight) or \"large\" (i.e. high molecular weight). The same rules apply if we want to make separations more efficient or detection limits lower. However, the different properties of larger molecules (e.g. lower diffusion coefficients, limited solubility) lead to some differences in practical implementations of separation methods. Moreover, many different molecular structures exist, both for synthetic polymers (exhibiting molecular-weight distributions, chemical-composition distributions, etc.) and for biomolecules, such as proteins, which show many different proteoforms.<br \/>Retention of large molecules in LC depends very strongly on the mobile-phase composition. This makes isocratic analysis very difficult and, thus, gradient-elution LC the preferred method. However, gradients will only separate highly similar large molecules, such as proteoforms, if they are vary slow (i.e. span a small change in composition and\/or a large number of column volumes). Oppositely, fast gradients may be used to suppress the effect of molecular weight on retention of synthetic polymers, resulting in \u201cpseudo-critical\u201d separations based on chemical composition or end groups.<br \/>Sized-based separations, such as size-exclusion chromatography, hydrodynamic chromatography and field-flow fractionation, provide a different kind of selectivity, ideally suppressing any effects of composition on retention. <br \/>The existence of very different (\u201corthogonal\u201d) selectivities opens the door to highly informative multidimensional separations, with applications in purity testing or purity enhancement of target molecules, or comprehensive characterization of complex samples.<br \/>Examples will be discussed to explain the above concepts in this tutorial.<\/p><p>- - -<\/p><p><strong>Peter Schoenmakers<\/strong> has been a full-time professor in Analytical Chemistry at the University of Amsterdam since 2002 \u2013 with emeritus status since 2021. His has worked on many aspects of one- and two-dimensional chromatography, with a special interest in the characterization of macromolecules. He has seen method-optimization and artificial-intelligence tools emerge some forty years ago, with only gradual progress until recently. Finally, we are starting to see exciting progress in method development for LC.<\/p><p>Together with Bob Pirok, Peter Schoenmakers has recently completed a new textbook on Analytical Separation Science, which is to be launched at HPLC2025 Bruges. He is also the Education Director of COAST, The Netherlands\u2019 public-private-partnership organization on analytical chemistry. \u00a0From 2005 to 2020 he was an editor of the Journal of Chromatography A. He has received a number of international awards, most recently the ACS Award for Chromatography (2022) and the LCGC lifetime-achievement award (2023).<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Tutorial Description <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-805f8da e-flex e-con-boxed e-con e-parent\" data-id=\"805f8da\" data-element_type=\"container\" data-settings=\"{&quot;background_background&quot;:&quot;classic&quot;}\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t<div class=\"elementor-element elementor-element-fc5e589 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"fc5e589\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-4085a0d e-con-full e-flex e-con e-parent\" data-id=\"4085a0d\" data-element_type=\"container\">\n\t\t<div class=\"elementor-element elementor-element-addf0b5 e-grid e-con-full e-con e-child\" data-id=\"addf0b5\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-54159f7 elementor-widget elementor-widget-image\" data-id=\"54159f7\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" src=\"https:\/\/hplc2025-bruges.org\/wp-content\/uploads\/elementor\/thumbs\/Mark-Schure-2-qzy36548vk68d7ud89yazzfjviro1v6ec90v72h5ns.jpg\" title=\"Mark Schure 2\" alt=\"Mark Schure 2\" loading=\"lazy\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-7c1a3d0 elementor-widget elementor-widget-text-editor\" data-id=\"7c1a3d0\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00a0 \u00a0<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-2777be1 e-con-full e-flex e-con e-child\" data-id=\"2777be1\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-71a92e5 elementor-widget elementor-widget-text-editor\" data-id=\"71a92e5\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>TUTORIAL 06 &#8211; Thu June 19 &#8211; 10h45 \/ 11h30<\/p><p>MARK <strong>SCHURE<\/strong><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-6619d86 elementor-widget elementor-widget-heading\" data-id=\"6619d86\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">Molecular simulations, molecular measurements, how they work and what we can learn for chromatography.<\/h2>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-07b7d82 e-con-full e-flex e-con e-child\" data-id=\"07b7d82\" data-element_type=\"container\">\n\t\t\t\t<div class=\"elementor-element elementor-element-00aae8c elementor-widget elementor-widget-tp-unfold\" data-id=\"00aae8c\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e065827 tp-ca-center \" data-id=\"plus6a9679e065827\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Speaker Biosketch\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p><strong>Mark Schure<\/strong>\u2019s scientific interests include fundamental separation science, the separation of polymers and colloids, and all aspects of solving large-scale chemical and physical problems with computers. His contributions to separation science include detailed theory, simulations and experimental investigations in the areas of 2D chromatography, chromatographic stationary phase retention mechanisms, capillary electrophoresis, electrochromatography and field-flow fractionation.<\/p><p>He has received the Arthur Doolittle award from the PMSE (Polymeric Materials Science and Engineering) Division of the ACS, the Northeastern University Distinguished Alumni Lecture award, the Douglas Leng award within The Dow Chemical Company, the L. S. Palmer award from the Minnesota Chromatography Forum, the Eastern Analytical Symposium award in separation science, the Stephen Dal Nogare award and the Uwe D. Neue award. Dr. Schure has been an Adjunct Professor of Chemical Engineering at the University of Delaware for over 30 years and has published over 120 papers and 4 patents.<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Speaker Biosketch <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-6bbfc8d elementor-widget elementor-widget-tp-unfold\" data-id=\"6bbfc8d\" data-element_type=\"widget\" data-widget_type=\"tp-unfold.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<div class=\"tp-unfold-wrapper plus6a9679e065d69 tp-ca-center \" data-id=\"plus6a9679e065d69\"  data-content_a_source=\"content\" data-con_pos=\"default\" data-icon-position=\"tp_ic_pos_before\" data-readmore=\"Tutorial Description\" data-readless=\"Read Less\" data-readmore-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book-open&quot;&gt;&lt;\/i&gt;\" data-readless-icon=\"&lt;i aria-hidden=&quot;true&quot; class=&quot;fas fa-book&quot;&gt;&lt;\/i&gt;\"  data-transition-duration=\"200\" data-content-max-height=\"100\" data-content-max-height-t=\"100\" data-content-max-height-m=\"100\"  ><div class=\"tp-unfold-title\"><\/div><div class=\"tp-unfold-description\" ><div class=\"tp-unfold-description-inner\"><p>Molecular simulations offer a unique view of chemical processes that experiment and theoretical approaches can not give. These techniques include Monte Carlo, molecular dynamics, and simple computer methods for measuring distances, interactions, and where solutes interact with the chromatographic phases, once atom-based models are created. Molecular simulations will be described which have been applied to all forms of liquid chromatography, from reversed-phase (RPLC), hydrophilic interaction (HILIC) to supercritical fluid chromatography (SFC) and will contrast these techniques in terms of phase interactions, geometry, and the thermodynamic differences in solute interactions. This tutorial is intended to reveal the physics which underlie these techniques, the chromatographic differences and what the atomic picture teaches us about these forms of chromatography.\u00a0<\/p><\/div><\/div><div class=\"tp-unfold-last-toggle center\"><button class=\"tp-unfold-toggle \"><i aria-hidden=\"true\" class=\"fas fa-book-open\"><\/i> Tutorial Description <\/button><\/div><\/div>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-518b102 e-flex e-con-boxed e-con e-parent\" data-id=\"518b102\" data-element_type=\"container\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t<div class=\"elementor-element elementor-element-ad47ff9 elementor-widget elementor-widget-spacer\" data-id=\"ad47ff9\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<div class=\"elementor-element elementor-element-63d9f13 e-flex e-con-boxed e-con e-parent\" data-id=\"63d9f13\" data-element_type=\"container\">\n\t\t\t\t\t<div class=\"e-con-inner\">\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t","protected":false},"excerpt":{"rendered":"<p>Tutorials \u00a0 \u00a0 TUTORIAL 01 &#8211; Wed June 18 &#8211;\u00a0 08h30 \/ 09h15 LARS NILSSON Asymmetric flow field-flow fractionation (AF4) coupled to synchrotron small angle x-ray scattering (SAXS) \u2013 A new tool for characterization of proteins and nanoparticles Lars Nilsson has a background in surface and colloid science, receiving his PhD from Lund University, Sweden&#8230;<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"elementor_header_footer","meta":{"footnotes":""},"class_list":["post-1069","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=\/wp\/v2\/pages\/1069","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1069"}],"version-history":[{"count":52,"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=\/wp\/v2\/pages\/1069\/revisions"}],"predecessor-version":[{"id":2217,"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=\/wp\/v2\/pages\/1069\/revisions\/2217"}],"wp:attachment":[{"href":"https:\/\/hplc2025-bruges.org\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1069"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}