RNA-based products are revolutionizing the field of advanced therapies, including the development of prophylactic vaccines, cancer vaccines, and genome-editing products. In this presentation, I briefly introduce various product types under investigation and provide guidance on the quality of materials used in the manufacturing of different product classes.

The quality and purity of mRNA are critical for vaccine and therapeutic safety. As new technologies emerge, bridging analytical methods, especially in impurity detection, remains challenging. Comprehensive characterization is essential for identifying impurities like dsRNA and residual DNA. USP is developing documentary and physical standards which support the mRNA product lifecycle. This presentation explores evolving mRNA analytics and opportunities to enhance impurity detection and improve product reliability.

This presentation will delve into the analytical testing methodologies critical for the regulatory CMC submissions of self-amplifying mRNA therapies. It will cover the development of robust analytical strategies to assess the quality, potency, and stability of these mRNA constructs. Key challenges such as assay validation, sensitivity optimization, and meeting regulatory standards will be discussed, alongside case studies highlighting successful CMC documentation for approval processes.

The emergence of mRNA modality as a therapeutic alternative for treatment of viruses, infectious disease, and various therapies has highlighted the need for high quality mRNA. To meet the quality requirement, developers continue to examine raw material quality, process controls and appropriate analytical methods to ensure the manufacturability of high quality mRNA. As such, we will discuss various strategies to ensure the manufacturability and characterization of high-quality mRNA.

Gene editing therapeutics present unique CMC challenges, from raw material sourcing to scalable manufacturing and cost optimization. As these therapies progress toward the clinic, ensuring process and analytical robustness, and economic feasibility is critical. This talk will explore key CMC considerations in gene editing, comparing its manufacturing and cost dynamics to oligonucleotide-based therapies. Key topics include manufacturing challenges for pDNA, mRNA, and LNPs, and analytical hurdles in mRNA, gRNA, and LNP characterization. Strategies for outsourcing, tech transfer, and CDMO engagement will be explored, with a focus on integrating innovative technologies with practical manufacturing solutions.

mRNA-based therapeutics are advancing, but downstream processing remains challenging due to low yields and high costs. We present an integrated, continuous manufacturing process for mRNA purification using precipitation-based methods, followed by continuous flow filtration. In these processes, the 3D structure of precipitates is crucial for recovery and dissolution. To address this, we introduce an AI-driven high-throughput image analysis system to screen and characterize precipitates and precipitation conditions, optimizing filterability.

Current RNA manufacturing generates dsRNA impurities that must be removed, along with enzyme(s) and DNA, in purification. Functional co-immobilization of enzyme and DNA to a solid support prevents the formation of dsRNA, and eliminates costly purification. This allows a single-use chip, continuous flow reactor for a single-path workflow from NTPs to highly pure RNA of any length. New analytics allow for real-time quality and yield optimizations in long continuous production runs at all scales.

• Examine the evolving purification challenges for RNA and oligonucleotide therapeutics, including impurity profiles and product heterogeneity
• Compare platform strategies versus molecule-specific solutions in downstream development.
• Highlight key technologies—such as chromatography, TFF, and enzymatic treatments—advancing purification efficiency and scalability
• Explore regulatory expectations and process validation considerations for RNA and oligo manufacturing

With the growing importance of ASOs, synthetic chemists are increasingly focused on developing more efficient and environmentally sustainable synthesis methods. Biogen has developed a liquid-phase-oligonucleotide-synthesis (LPOS) process to increase the synthesis output while maintaining product quality. However, LPOS has long development time compared to solid phase synthesis (SPS). We recently developed a Hybrid Synthesis Platform utilizing a novel resin developed in house. This resin enables rapid preparation of protected oligonucleotide fragments via SPS followed by LPOS assembly to full length product. The hybrid synthesis approach significantly shortens LPOS production cycles, making it suitable for supporting early-stage clinical studies.

In vitro transcription is an RNA synthesis step used in non-viral gene therapy, including mRNA or CRISPR guide RNA, where linearized DNA serves as a template for the enzymatic reaction. Conventional methods for preparing the DNA template have required large-scale microbial cultivation, plasmid purification, and linearization using additional restriction enzymes. In this talk, we present a novel method for DNA preparation using continuous, programmable PCR amplification enabled by flow chemistry. This research was supported by the US Food and Drug Administration under the FDA BAA-22-00123 program, Award Number 75F40122C00200.

This presentation will address the unique chemistry, manufacturing, and controls (CMC) challenges associated with prime-edited therapies. It will explore the intricacies of developing and scaling up prime editing platforms, focusing on aspects such as product design, delivery systems, and manufacturing processes. The session will also discuss regulatory hurdles, strategies for ensuring product consistency and purity, and the critical role of CMC in advancing prime-edited therapies towards clinical application.

Gene therapy has re-emerged as a new therapeutic modality due to the recent breakthroughs and the potential to provide a long-term treatment for a genetic disease for patients. Traditionally, viral vectors, such as adeno-associated virus (AAV), have been widely used but pose challenges such as potential humoral immune response, limitation of re-dose, and limited cargo capacity. In contrast, non-viral delivery with lipid nanoparticles (LNPs) can carry larger genetic payloads (> 10kb for DNA in theory) and the ability to re-dose the patients once the treatment wears off. This presentation will explore the latest advancements in the analytical methods used to evaluate lipid nanoparticles (LNPs). It will cover a range of cutting-edge techniques that improve the characterization and understanding of LNPs, focusing on their application in drug delivery systems. Key topics will include the development of more precise and efficient analytical tools, their impact on quality control, and their role in enhancing the overall development process of LNPs.

This presentation will discuss the role of digital technologies in enhancing the development of lipid nanoparticle (LNP) formulations. It will focus on the integration of computational tools, data analytics, and machine learning to refine LNP design and manufacturing processes. It will examine how these technologies contribute to improved efficiency, scalability, and precision, ultimately facilitating faster and more reliable production cycles for advanced therapeutic delivery systems.

Lipid nanoparticles (LNPs) are a versatile and effective platform for delivering nucleic acid therapeutics, as demonstrated by the COVID-19 vaccines. Despite a conceptually straightforward manufacturing process, challenges in understanding LNP formation and scale-up persist. This presentation outlines a multiscale mechanistic modeling framework, leveraging thermodynamic and phase-field models to predict properties, study RNA encapsulation, and optimize processes. Links between the model(s) and product quality, process control, and optimization are discussed.