Towards the high throughput production of oligonucleotides – Insights into upstream, downstream and analysis
Thermo Scientific: Towards the high throughput production of oligonucleotides – Insights into upstream, downstream and analysis
Besides being the major provider for genotyping and DNA sequencing services, Eurofins Genomics Europe Pharma and Diagnostic Products & Services Synthesis GmbH is one of the world-leading oligonucleotide manufacturers for the industrial and research sector.
Since 2021, Eurofins Genomics can capitalise of next generation high throughput synthesizers, the so-called Continuous Flow Synthesizer (CFS), which allow a simultaneous production of 6 x 384 different oligonucleotides with elevated speed reaching an excellent coupling efficiency of up to 99.6%.
We present an elaborated upstream and downstream process for the high throughput production of standard and modified oligonucleotides of a length up to 200 nucleotides, including insights into the chemical background. Beside the synthesis procedure, the sample purification and analysis are the major hurdles in oligonucleotide production. The Thermo Scientific™ Vanquish™ IP-RP-UPLC, commonly used with customary C18 columns to obtain highly resolved analytical chromatograms for an unambiguous sample composition determination, was proven to be very suitable for semi-preparative purification applications for oligonucleotides like dual-labeled probes. In addition, we showcase the crucial role of LC-ESI-MS investigation on a modified DNA longmer leading to an optimized production strategy and hence resulting in a significantly improved impurity profile which massively simplifies the subsequent chromatographic purification.
Learning points:
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Discover an upstream and downstream process for the high throughput production of standard and modified oligonucleotides of a length up to 200 nucleotides.
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Learn how to overcome challenges regarding sample purification and analysis.
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Learn about the crucial role of LC-ESI-MS investigation on a modified DNA longmer leading to an optimized production strategy
Who should attend:
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Biopharmaceutical Research Scientists
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Analytical Scientists from Biopharma companies working on or interested in oligonucleotide-based therapies.
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PhD students in any field of biotherapeutic drug development
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Scientists looking to advance knowledge of oligonucleotide analysis.
Presenter: Dr. Johannes Kremser (Senior Scientist R&D for Oligonucleotides, Eurofins Genomics)
With more than seven years of experience in the field of oligonucleotides, Dr. Johannes Kremser has a proven track record of successful project management and producing quality outcomes through professional expertise and team motivation. His advanced education began in Innsbruck at the Leopold-Franzens University where his bachelor thesis in Organic Chemistry was supervised by Univ. Prof. Ronald Micura. Upon completion of his bachelor’s degree in 2014, he continued there under the direction of Assoc. Prof. Christoph Kreutz where he defended his masters in Organic Chemistry in 2016.
His passion for oligonucleotide chemistry in combination with NMR spectroscopy took him abroad to London as a visiting doctoral student at the University College London where in the Department of Structural and Molecular Biology guided by Dr. D. Flemming Hansen, his project focused on RNA entitled ‘Elucidation of transiently formed excited state RNA structures by NMR spectroscopy’. With his successes in the UK, he returned in 2019 to his alma mater of Leopold-Franzens University to produce and defend his doctoral thesis “'Seeing the Invisible' - Investigation of RNA Excited States via NMR Spectroscopy” under supervision of Assoc. Prof. Christoph Kreutz.
Johannes transitioned from his academic career into his professional debut as the group leader in R&D for oligonucleotides in Switzerland at Bachem AG. His subsequent and present employment with Eurofins Genomics in Ebersberg commenced in 2022 as Senior Scientist R&D for Oligonucleotides where he manages the laboratory and process planning for RNA high throughput and mid-scale production and ensure ISO-conform oligonucleotide production.
Presenter: Dennis Köhler (Product Application Specialist, Thermo Fisher Scientific)
Dennis Köhler has a pronounced career ranging from organic synthesis in the field of oligonucleotides as well as analytics for almost 2 decades. Graduating from the University of California, San Diego in 2004 he immediately initiated his professional path in the region of modified nTP synthesis and purification at Trilink Biotechnologies, Inc. Subsequently, he spent short of 10 years synthesizing fluorophores for DNA labelled FRET technologies for Biosearch Technologies, Inc. After moving to Vienna in 2014, he joined the Thermo Scientific team as sales support for chromatography where in 2018 he transitioned to the Thermo Scientific Product Applications team outside of Munich, contributing to application notes, collaborations, new product development, and elevated field support.