Synthetic oligonucleotides are being employed as therapeutics with multiple mechanisms of action. A variety of mechanisms leverage hybridization to RNA to modulate the translation of desired proteins. Separate oligonucleotides have also been designed to interact with pattern recognition receptors, leading to the modulation of an immune cascade. Though therapeutic when intended, this immune modulation must be accounted for as unintended immune activity could also impact the safety of these therapeutics.

Speaker:

Sudhir Agrawal, D.Phil
Founder & President
Arnay Sciences

Anion exchange chromatography as a purification technique for oligonucleotides will be discussed. We will present different case studies where we have purified DNA and RNA oligomers, as well as a pegylated RNA, how we have optimized the purification parameters and challenges that may occur.

Speakers:

Cecilia Unoson
Manager Applications
Bio-Works

Over the last two decades, DNA synthesis has established itself as a standard tool propelling life science research in a wide range of fields. In this session, we will highlight how DNA synthesis tools can support mRNA therapeutics development.

One key benefit of synthetic DNA is that it enables flexible sequence design independent of any naturally occurring template. Methods for optimization of expression in a target organism will be discussed, focusing on how to achieve best results in mammalian systems. The ability to design sequences also enables incorporation of additional layers of information into the DNA. To this end, a method for “molecular watermarking” will be described, which can enable tracking and tracing of a molecule throughout the application.

mRNA Therapeutics have a wide range of applications ranging from personalized approaches in cancer immunotherapy to global applications in the infectious disease landscape. Depending on the application, the needs for DNA input differ widely. These needs will be discussed using specific examples of how Thermo Fisher GeneArt DNA synthesis technologies can address a variety of needs across myriad applications in the life science industry.

In summary, this presentation will highlight how different Synthetic Biology technologies can support emerging needs in mRNA therapeutics development and commercialization.

Speaker:

Axel Trefzer
R&D leader
Thermo Fisher Scientific

Oligonucleotide and mRNA drug products or vaccines are specific in their mode of action and are typically administered via sub-cutaneous or intravenous routes. However, the potential for administration through inhalation has been steadily growing in importance as this presents a more desirable mode of self-administration, resulting in an improved patient experience and compliance. Therefore, if viable, inhalation offers a highly attractive non-invasive route for the administration of various classes of oligonucleotides, not only for the treatment of respiratory diseases through targeted delivery to the lungs, but also as an alternative route to parenteral administration.

In this presentation we cover key considerations for inhaled or nasal delivery including formulation, device selection / compatibility and analytical development from both the molecule (structure, physicochemical properties, potency) and delivery performance aspects (delivered dose, delivery rate, aerodynamic droplet size distribution, etc.). We will also include a case study involving the development of a nebulized oligonucleotide.

Speaker:

Ashleigh Wake
Director of Business Development C&P UK
Intertek

Genetically-encoded (GE) libraries of peptides are a well-established and readily available technology. “Late stage” modification of these GE libraries in water, when optimized, can at once convert million to billion diverse starting materials to products with drug-like properties. My talk will focus on recent transformations of GE-libraries of peptides to yield GE-libraries of novel macrocyclic architectures with unnatural pharmacophores. I will show that the resulting libraries accelerate ligands discovery in early drug discovery. These libraries are ideally poised for fragment-based discovery and optimization of specificity and potency of the existing fragments / pharmacophores.

Speaker:

Ratmir Derda, Ph.D.
Associate Professor, Department of Chemistry
University of Alberta, Canada

Peptide purification challenges can be solved through optimization of the synthesis reaction or by applying downstream purification technology. In this panel discussion, experts will discuss the advantages and disadvantages to both strategies and answer audience questions.

Moderator:

Jonathan Royce
CEO
Bio-Works

Panelists:

Richard E. Johnsson, Ph. D., Associate Professor
Head of Peptide Chemistry
Red Glead Discovery

Elizabeth Denton, PhD
Senior Scientist, Peptide Chemistry
Biotage LLC

Sanjay D. Patil
Sr. Manager- Peptide Production
Sun Pharma

Development of methods aimed at increasing sustainability of peptide synthesis and purification has experienced enormous interest in recent years as evidenced by the numerous publications and reviews disclosed in the area. Nevertheless, the uptake of these greener methods by practitioners has been lacking, with majority of reports on target-oriented peptide synthesis still relying heavily on hazardous starting materials and use of energy intensive methods and large amounts of process aids. Herein, we will discuss various aspects that impact the adoption of greener methods in synthetic peptide chemistry, illustrated by case stories of methods developed in our laboratories.

Speaker:

Jan Pawlas, Ph.D.,
Scientist in Global Development
PolyPeptide Group

Collagen is one of the fundamental components of the corneal extracellular matrix and a common base material for tissue regeneration. Animal derived collagens present challenges in terms of potential adverse responses to xenogenic material and the potential for disease transmission. Recombinant collagens are expensive to produce and limited in supply for material-based solutions. All atelocollagen requires manufacturing at 2-8 °C to maintain its fibrillar assembly and prevent conversion into gelatin due to heat and friction. Short collagen mimetic peptides have been designed to undergo self-assembly into fibrils and can substitute for atelocollagen in biomaterial solutions for regenerative medicine. They have the advantage of convenient and cost-effective solid-state synthesis, high temperature manufacturing process to promote fibrillar assembly, and no risk of patient exposure to xenogenic material. Here we discuss the use of CMPs in both solid corneal implants and a LiQD Cornea that acts as a corneal sealant for the regeneration of corneal tissue in situ.

Key Takeaways:

• Self-assembling peptides can substitute for atelocollagen in hydrogel-based biomaterials.
• Peptide-based materials allow for customized functionalization for conjugation.
• Peptides can reduce manufacturing costs for biomaterial production.

Speaker:

Fiona Simpson
PhD Candidate
Université de Montréal & Hôpital Maisonneuve-Rosemont Research Center

Site-selective modification of proteins is highly desirable for the introduction of small probes or larger moieties such as PEG. In 2018 we reported the development of a Gly-His3-6 tag for selective N-terminal acylation of proteins.1 Here, we report new reagents and mechanistic studies. Next, we extended His tag acylation by developing a new autocatalytic His sequence for highly selective acylation on Lys, which we name a Lys-His tag. This provided selective chemical modification of antibodies.

Speaker:

Knud Jørgen Jensen, Ph.D.
Professor, Department of Chemistry
University of Copenhagen

The application and reliance on Solid Phase Peptide Synthesis (SPPS) has increased dramatically over recent years with the rise of peptides in diagnostic and therapeutic research. With this rapid increase in development activity, automated synthesis methods have had to adapt to be compatible with the wide range of coupling chemistries, reaction conditions and synthesis monitoring available to the practicing chemist. The subsequent purification of the synthesized peptides in that process demands rapid scale-up what is easily achieved using the Peptide Easy Clean (PEC) technology. The need for parallel, simultaneous reaction screening to optimize SPPS and purification conditions has never been greater. Here we describe the automated screening of multiple reaction conditions for the synthesis of a selection of therapeutic peptides, enabling the optimum synthetic conditions to be selected for each target, followed by the scale-up of the synthesis on a pilot-scale instrument. The rapid purification from mg to gram-scale using the PEC technology is show-cased on Liraglutide.

Speaker:

Andrew Kennedy, Ph.D.
Global Product Manager (Peptides Business)
Gyros Protein Technologies

Dr. Robert Zitterbart
Co-Founder, Head of R&D
Belyntic GmbH, Berlin

The primary method for quantitation and characterization of synthetic oligonucleotides is liquid chromatography-mass spectrometry (LC-MS). Whether for characterizing process and product-related impurities or bioanalytical pharmacokinetic studies, improving method sensitivity is of critical importance. In this presentation, we discuss how LC-MS sensitivity can be improved with mobile phase optimization and column selection. We also demonstrate how one can minimize adduct formation and improve ionization efficiency, which can improve MS sensitivity for improved characterization and quantitation data.

Speaker:

Brian Rivera
Senior Product Manager – Biologics
Phenomenex

SCIEX recognizes that in-depth characterization and precise quantification of peptides and oligonucleotides such as RNAs or antisense oligonucleotides (ASOs) is challenging. We aim to make you more productive with end-to-end CE and LC-MS solutions. In this workshop we will cover both new characterization approaches for RNA therapeutics from small to large, as well as sub-ng/mL quantification of ASOs in rat plasma using trap-and-elute microflow LC-MS/MS. The latter is of particular interest for studies where ASO sample is limited, e.g., metabolism studies in rats or mice, and sensitivity may still be insufficient. During these two presentations, you will understand where these techniques fit into your analytical toolbox so you can:

• Detect and highly resolve RNA molecules.
• Speed up RNA aptamer research.
• Achieve in-depth characterization of ASOs.
• Attain sub-ng/mL quantification of ASOs in plasma for low amount of sample volume for limited samples, increasing sensitivity by a factor of approximately 10x compared to previously published data acquired using standard LC flow rates.

Speakers:

Dr. Lei Xiong
Sr. Manager Biopharma Application Science
SCIEX 

Dr. Stephen Lock
Marketing and Market Development Manager
EMEAI 

• There are no well-defined guidelines for therapeutic oligonucleotide development by any authorities.
• Syngene has developed a platform technology to manufacture oligonucleotide drug substances using a systematic approach for process and analytical methodologies to address the issue.
• We will be presenting some process and analytical quality control challenges encountered during the development activities of oligonucleotide drug substances.

Speaker:

Dr. Chanchal K. Malik
Lead Investigator - Oligonucleotides
Syngene International Ltd  

Dr. Guru S. Madugundu
Lead Scientist - Oligo Analytical Development
Syngene International Ltd

The current modus operandi for the downstream processing of synthetic peptides is reversed phase chromatography performed in batch mode. Continuous chromatography is expected to be a disruptive technology for this very cost intensive part of synthetic peptide manufacturing. Results from a proof of principle study will be presented and compared to traditional batch chromatography.

Speaker:

Dr. Ralf Eisenhuth
Process Manager Chromatography and Technology Transfer
Bachem

Synthetic oligonucleotides are manufactured with a hydrophobic trityl protecting group that serves as the chromatographic handle for purification. When the trityl group is no longer needed, there are two deprotection schemes widely used in the oligonucleotide industry: anion exchange chromatography with on-column deprotection (AEX-OCD) or solution-phase deprotection. The AEX-OCD route exposes the oligonucleotide to both acidic and basic conditions which can lead to product degradation. The solution-phase deprotection route involves several ethanol precipitations steps that can be very time consuming.

A new manufacturing process was developed where solution- phase deprotection is performed in a TFF skid retentate vessel. This new process results in less potential for oligonucleotide degradation compared to AEX-OCD and requires less processing time compared to traditional solution-phase deprotection process involving ethanol precipitations.

Speaker:

Mr. Dana Chreng
Associate Director- Drug Substance Manufacturing Group
Ionis Pharmaceuticals

The application of Twin-column continuous countercurrent chromatography (the “MCSGP” process) for Oligonucleotide purification will be shown in this presentation. MCSGP simultaneously achieves high yield and purity through automatic internal recycling of impure side fractions. Thereby MCSGP avoids excessive peak fractionation, fraction analysis and re-chromatography. Consequently, in production scale, much fewer samples need to be analyzed by QC and release times are shortened, allowing faster project turnover. This talk briefly introduces the MCSGP process concept and recent results of oligonucleotide purification by countercurrent chromatography and presents the key economic aspects for scale-up.

Speaker:

Thomas Müller-Späth, Ph.D,
Chief Technical Officer
YMC ChromaCon AG, Switzerland

Insights by a panel of industry thought leaders.

Moderator

J Preston Ph.D.
Director, Separations Science and a Senior Subject Matter Expert
YMC America

Panelists

Dr. Ralf Eisenhuth
Process Manager Chromatography and Technology Transfer
Bachem

Mr. Dana Chreng
Associate Director- Drug Substance Manufacturing Group
Ionis Pharmaceuticals

Thomas Müller-Späth, Ph.D,
Chief Technical Officer
YMC ChromaCon AG, Switzerland

ApTOLL is an aptamer designed to block TLR4, a crucial receptor involved in the inflammatory response triggered in different diseases. Efficacy and safety of ApTOLL have been demonstrated in experimental models and replicated in different animal species and different laboratories blinded to the treatment. Additionally, a First-in-Human clinical trial has been completed with exceptionally good results in healthy volunteers and a Phase Ib/IIa is currently being conducted in stroke patients.

Speaker:

Macarena Hernández-Jiménez, Ph.D.
Chief Scientific Officer
AptaTargets S.L.  

Description Coming Soon

Speaker:

Huihe (Julia) ZHU, Ph.D.
Senior Manager
Small Scale Oligonucleotide Unit
Nitto Avecia  

As technologies for unbiased library screening, incorporating both natural and non-natural amino acids, have evolved, the appetite for pursuing discovery projects with peptidic compounds has increased. Automated parallel peptide synthesizers enable the synthesis of hundreds to thousands of compounds simultaneously, that then feed into the lead optimization campaign. However, there are very few solutions that address peptide purification, or even a simple clean up, in a high-throughput manner. Herein, we introduced an automated strategy to improve the purity of crude synthetic peptides in parallel.

Speaker:

Dr. Elizabeth Denton
Senior Peptide Scientist
Biotage