Mark Hilliard’s Post

🧬 Tuning RNA Digests for Seq+Mod Mapping #mRNA and #CRISPR sgRNA drug substances are on the move. The analytical labs supporting them have been hard at work confirming their sequences, double checking impurities, and measuring their important modifications - all of which are tied to their safety and efficacy. These aren’t the easiest analyses to perform. 💡We think bottom-up analysis of #RNA therapeutics should be just as straightforward and accessible as peptide mapping and proteomics experiments. So, our teams and our collaborators are on the move, too. 👇We just finished a major project on new RNase specificities for sgRNA and mRNA sequence and modification mapping. An example of an improved uridine-specific digestion is shown below. With a new level of partial digestion tunability and a choice of 5' uridine and 3' cytidine specificities, each and every detail of a large RNA therapeutic can be tracked and magnified for in-depth, comprehensive physicochemical analyses. 💻 Stay tuned this Thursday for in-depth announcements and join QUALITY ASSISTANCE S.A. and Waters Corporation scientists for our "Cracking the Code" webinar on Oct 16th! Webinar Link: https://lnkd.in/eAsJDZQq Waters.com/RNA #GeneTherapy, #CellTherapy, #CMC, #AnalyticalChemistry, #LCMS, #HPLC, #MassSpec #LifeatWaters #biotechnology

𝗨𝗻𝗱𝗲𝗿𝘀𝘁𝗮𝗻𝗱𝗶𝗻𝗴 𝘁𝗵𝗲 𝗥𝗼𝗹𝗲 𝗼𝗳 𝗟𝗶𝗽𝗶𝗱𝘀 𝗶𝗻 𝗠𝗲𝗺𝗯𝗿𝗮𝗻𝗲 𝗣𝗿𝗼𝘁𝗲𝗶𝗻 𝗙𝘂𝗻𝗰𝘁𝗶𝗼𝗻 Lipids are not just structural components of cellular membranes; they play crucial roles in the function of membrane proteins. This dynamic interplay between lipids and proteins is fundamental to molecular cell biology. 🔬 Recent Findings: • Lipids act as specific cofactors and solvents for membrane proteins, affecting their activity and stability. •The diversity of lipids in mammalian cells suggests a nuanced regulatory layer for IMPs, which is influenced by cell type, metabolic state, disease state, and external factors like diet. 📚 Why It Matters: • Understanding these interactions opens new avenues for biomedical research, particularly in drug targeting, as the protein-lipid interface is a key site for pharmaceutical intervention. • The concept of the 'functional paralipidome' highlights how specific lipid environments can regulate IMP functionality, providing a framework for future studies. The paper by Ilya Levental and Ed Lyman offers a comprehensive review of these interactions and their implications for cellular function. It's a must-read for anyone interested in the intricate dance of lipids and proteins in cellular membranes. https://lnkd.in/ecm-_zya #MolecularBiology #CellBiology #Biochemistry #MembraneProteins #Lipidomics #BiomedicalResearch #Science

💊Changing the focus to solve a familiar challenge: dsRNA impurities. dsRNA populations are common impurities in mRNA production that appear during IVT process. These contaminants can trigger immune responses or induce off-target effects, affecting the safety and efficacy of the therapeutic agent. 🔍Origins of dsRNA are not crystal clear yet. Most theories focus the responsibility of dsRNA generation on the enzyme in charge of the nucleotide polymerization (T7 RNA polymerase). For that reason, efforts to avoid and remove these impurities are commonly concentrate in post-IVT purification. Therefore, rigorous purification methods and quality control measures are necessary to minimize dsRNA contamination. Typically, two chromatographic steps are used for that purpose: affinity + anion exchange chromatography. Researchers of RNA department of Certest Pharma have changed the focus to provide a new starting point to reduce dsRNA presence. During their research, they noticed that different integrity, purity, and isoform profile of linear DNA generated from plasmids and used as templates for IVT could affect to dsRNA generation. So, they decide to check different chromatographic techniques to purify linear templates and study how dsRNA was affected. Therefore, they have optimized hydrophobic interaction chromatography purification of linear templates resulting in the decreasing of dsRNA during IVT. This research opens the gates to a new approach for the reduction of dsRNA content and suggest a new mechanism of generation of this important impurity. Are you interested in knowing more about this work? 🔗Click here to download the paper. https://lnkd.in/d9vkeX5v #CertestPharma

Some very interesting work by Zao et al looking g at the effect on improving MS/MS sequencing of Oligonucleotides. 🛎 Overview: Zao et al out lines that negative-ion mode fragmentation such as CID or HCD are impacted by their terminal phosphate in an unwanted manner.  This is believed to be mediated via two ways. 1) the terminal phosphate product an increased negative chage state 2) The terminal phosphate tends to be lost in MS2 spectra. This loss results in the decrease intensity of sequencing ions and thus impacting over sequencing coverage. 🎯 Summary: Here Zao et al rather cleverly employs calf intestinal alkaline phosphates (CIP) to remove the terminal phosphate group. Zao et al observed that with CIP treatment there was improved sequence coverage of oligonucleotides with terminal phosphates. #massspectrometry #massspec #science #sciencecommunication #chemistry #biotech #biopharmaceuticals #mrna #dna #oligonucleotides #nucleosides #biotechnology #science #sciencecommunication #scienceandtechnology #nucleotides https://lnkd.in/eYJ_DxjH 

Recently, there has been a growing interest in utilizing #ComputationalTools to redesign #mRNA sequences to improve #vaccine characteristics and expedite discovery timelines. In this review, they explore important biophysical features of mRNA to be considered for vaccine design and discuss how #computational approaches can be applied to rapidly design mRNA sequences with desirable characteristics. https://lnkd.in/gDivyn3T #mRNA #AI #SyntheticBiology #RNA Jiangsu Synthgene Biotechnology Co., Ltd.-mRNA Raw Material,Phosphoramidites,Molecular Diagnostic Raw Material,Customized service (synthgene-bio.com)

#TechnoteTuesday -Exciting Advances in Drug Development! Circular RNA Takes Center Stage In the world of cutting-edge drug development, Circular RNA (circRNA) is stealing the spotlight! Its covalently closed loop structure provides exceptional stability and boasts resistance to RNases, making it a promising therapeutic target. Researchers are making strides with a revolutionary analytical method that separates circRNA from its linear precursor. The key? RNA co-analysis with simplified sample preparation unlocks a high-resolution circRNA species separation. What's the magic behind it? Capillary gel electrophoresis with a laser-induced fluorescence detector (CGE-LIF)! This powerful combo detects circRNA and spots complex species like RNA concatemers, low-level impurities, and fragmentation. Our latest technical note showcases the separation of two synthesized circRNA products from their linear precursors. The circRNA species displayed a delayed migration compared to their linear counterparts, highlighting the precision of this breakthrough approach. #CircularRNA #DrugDevelopment #Innovation #ScienceBreakthroughs #ResearchRevolution In today's Technote, Mario A Pulido, Peter Holper, Jane Luo, Tingting Li Paul Baker, and Sahana Mollah from team SCIEX look at the "Separation of circular RNA from the linear precursor by multi-capillary electrophoresis." #TechnoteTuesday #pegRNA #crisprcas9 #crispr #BioPhase8800 #RNA9000 #mabs #development #sciex #CapillaryElectrophoresis #SciexforScience #genetherapy #rnatherapeutics #PA800Plus #immunotherapy #clinicaltrials #safety #biopharmaceuticals

Happy to share our recent paper published today in RSC Chemical Science (accepted manuscript) showcasing one of the early research outputs from the mRNA display platform at Novo Nordisk. https://lnkd.in/dYH9kKhD Having received a PhD from the Lab of Prof Christian Heinis, I have had many chances to see the power of bicyclic peptide formats for the identification of hits with desirable properties. Yet very few published studies had demonstrated their superiority vs linear or monocyclic formats in head-to-head comparison experiments. This is what motivated the present work, where we used a simple and efficient mRNA display protocol to isolate functionally active bicyclic ligands to FGFR3c outperforming the other two formats in affinity and plasma stability. I would like to thank all the colleagues and collaborators who made this publication possible, in particular my former master Student Silvana Smilla Zurmuehl, Novo Nordisk Mentor Martin Muenzel and MetID expert Christian N. Cramer. By the way, if you would like to know more about what we do and join us in those efforts, we have 3 open positions open (technician, scientist and senior scientist) ! #mrnadisplay #bicyclicpeptides #cyclicpeptides #MyFirstChemSci #openpositions

Chemically induced proximity (CIP) is a rapidly evolving field that has sparked interest in both basic research and drug discovery. CIP involves using synthetic molecules to co-opt natural biological pathways, recruiting neosubstrates that are not normally encountered or enhancing the affinity of naturally occurring interactions. This field has led to the emergence of several induced proximity modalities that modulate both proteins and RNA inspired by PROTAC. Interesting review highlighting the potential impact of CIP on drug discovery:https://lnkd.in/efUAM6vT https://lnkd.in/eiABB9DD

Understanding the properties linked to LNP function is key for mRNA applications, so what characterization tools are available? Register to our FREE webinar👇 🗓September 24 🕐16:30, to find out! Secure your spot 👉https://lnkd.in/dT5B33jx Our nanoparticle tracking analysis (NTA) expert Agnieszka Siupa will be joined by University of Utrecht LNP researcher Mariona Estapé Senti to discuss the use of fluorescence labelling techniques to inform property-function relationships in mRNA-LNP delivery vectors. Mariona Estapé works as a Postdoctoral Researcher in the Nanospresso-NL project, which aims to develop technology for producing personalized nucleic acid therapeutics at the point of care. Here’s what you’ll learn: • How to create robust physicochemical profiles of size and concentration using dynamic light scattering and NTA 👩🔬 • How to detect mRNA in lipid nanoparticle vectors 💉 • How to optimize fluorescence labelling for reliable use 🔬 #LNP #MRNA #Pharma

🚀 New Blog Alert! 🚀 We're excited to share our latest blog post, "Challenges in Antisense Oligonucleotide (ASO) PK Bioanalysis." Dive into the complexities of ASO bioanalysis and gain insights on: 🔍 ASO's Structural Diversity 🔍 Issues with Sensitive Detection Methods 🔍 Complexities Linked to Matrix Effects Antisense oligonucleotides (ASOs) present unique bio-analysis challenges, particularly in pharmacokinetic (PK) studies. Our blog explores these issues in-depth, providing valuable knowledge for those navigating this complex field. If you're encountering any hurdles in ASO bioanalysis, especially in PK studies, we want to hear from you! Let’s solve these challenges together. 🖱️ Read the full blog post here: https://lnkd.in/e7kxWm2k 🖱️ #ASO #Bioanalysis #Pharmacokinetics #PKStudies #LifeSciences #Research #AntisenseOligonucleotides #Nanorobots