We present a novel method for producing mRNA-loaded lipid nanoparticles (LNPs) with independent control over size and morphology. By decoupling particle formation from mixing, and using tunable fusogenic conditions followed by kinetic arrest, the approach enables rational LNP design. Demonstrated in an impinging jet mixer, the method supports predictive modeling and quality control.

Recombinant adeno-associated virus (rAAV) manufacturing faces significant challenges in scalability and cost-effectiveness, limiting the widespread application of rAAV-based gene therapies. This presentation showcases how computational modeling is a crucial tool for addressing these challenges. A continuous process for rAAV production is designed through mechanistic modeling and process optimization. The digitally designed continuous rAAV manufacturing process is experimentally validated, achieving high-titer rAAV production for 25 days.

We present a hybrid continuous bioprocessing platform integrating high-intensity perfusion and hybrid continuous downstream processing. The process delivers >10-fold productivity gains (up to 8 g/L-day) with enhanced process/product control and improved product quality. Designed for ease of implementation, the modular platform reduces COGs, enables near “just-in-time” capacity, and incorporates risk mitigation across upstream and downstream. Successfully implemented at manufacturing scale, it demonstrates how process intensification can accelerate and de-risk biologics production.

Continuous biomanufacturing is transforming recombinant protein production, offering efficiency and scalability. We introduce enGenes-e^Xpress, a growth-decoupled E. coli system that is genetically stabilized and mitigates adaptive evolution in a two-stage chemostat. This process enables sustained, high-yield recombinant protein production while reducing operational footprint, CAPEX, and OPEX. Our platform outperforms traditional fed-batch methods, making continuous production viable for biopharma and industrial biotech. By integrating cost-effective solutions with process intensification, enGenes-e^Xpress paves the way for decentralized, scalable biomanufacturing.

We have developed an innovative antibody capturing technology that efficiently and continuously purifies antibody proteins without using a chromatography column. This system allows continuous capture of monoclonal antibodies directly from cell-containing culture fluid without clarification. The basic principle is that the affinity binding reaction is promoted by forcibly mixing the monoclonal antibody and Protein A resin in an in-line static mixer, and the antibody-bound resin is separated and collected from the culture fluid by a hydrocyclone. The monoclonal antibody is then collected by dissociating the antibody protein from the collected resin. The resin from which the antibody protein has been desorbed is regenerated by mixing it with a regeneration buffer in a static mixer after CIP. This series of operations is continuously circulated by connecting multiple units and used for capturing. Since this technology does not use a chromatography column, there is no need for a clarification process to separate solids from the culture fluid, and not only does it eliminate the need for centrifuges and depth filters that were previously used, but it also does not require a chromatography column, which eliminates the need for column packing and qualification operations. This shortens the monoclonal antibody manufacturing process and makes it possible to use single-use for all processes, leading to reduced capital investment.

This talk will examine strategies for enhancing bioprocess platforms through process intensification and Process Analytical Technology (PAT) implementation. We'll explore continuous processing approaches, advanced monitoring systems, and real-time quality control methods that maximize efficiency while maintaining product integrity.

Early process development is crucial for reducing environmental impact and pursuing sustainable, cost-effective bioprocessing. Traditional metrics like Process Mass Intensity (PMI) are inadequate for predicting energy use and CO2 emissions. A new approach uses early process information to build a digital factory in BioSolve Process to evaluate cost, building, and process energy, enabling accurate environmental impact predictions. This allows for easy rescaling and assessment of various technologies and process options. This revolutionary modeling approach establishes new green metrics and facilitates predictive modeling by Clinical Phase II for optimizing cost and sustainability. It enables complete manufacturing optimization before the process is fixed, a currently unavailable capability. A case study explores strategies for minimizing environmental impact and costs through process intensification, optimization of water usage, and HVAC improvements.

This presentation is focused on developing and applying models to evaluate energy usage in bioprocess facilities. We describe the key facility inputs that should be included. Model structure is reviewed. The role of facility equipment type, cleanroom classification and facility size are discussed. Factors that most influence energy usage are discussed. An end-to-end review of facility attributes and those that most contribute to energy usage are discussed.

“Green by Design” is a strategic imperative to improve environmental sustainability metrics of our manufacturing processes. This effort is particularly important for more complex biologics such as bispecific antibodies, which often have a higher process mass intensity (PMI) than traditional monoclonal antibodies. Here, we demonstrate a combined approach of implementing facility fit modeling and downstream process optimizations to improve process sustainability through decreased water and raw material requirements.

• What new analytics and monitoring tools are most increasing the amount of valuable process data?
• What AI/ML-empowered tools are being employed to improve bioprocess development and control?
• How is AI/ML involved in the third stage of process validation: continued process verification (CPV)?
• What are the unique aspects of applying GAMP 5 to AI-empowered PAT tools in a GxP environment?

• ​Decision criteria business case: market size and dose of the product
• Distributed versus centralized manufacturing
• Stability of product and manufacturing complexity
  

We have developed a self-recycling perfusion bioreactor that leads to lower cost of goods and reduced environmental impact compared to conventional perfusion manufacturing processes. After harvesting mAbs we process spent media in our waste separation device that utilizes ion concentration polarization to remove ammonia and lactate. Our process allows us to "regenerate" media by replacing depleted nutrients and recycling them into the bioreactor without accumulating inhibitory metabolic waste products.

A mechanistic model is presented for a continuous lyophilization process. The model considers state-of-the-art lyophilization technology, in which vials are suspended and move continuously through the process. The model predicts the evolution of critical process parameters, including product temperature, ice/water fraction, sublimation front position, and concentration of bound water. The model is applied to process design and optimization of continuous lyophilization. The model is made available in MATLAB and Julia as a software package called ContLyo, which can be used to guide the design and development of continuous lyophilization processes.

In this talk, we discuss the development of a comprehensive modeling strategy for a continuous downstream process for monoclonal and multi-specific antibodies. The modeling framework includes (i) an end-to-end systems model for process design, optimization, and what-if analysis; (ii) residence-time distribution models to support control strategy definition; (iii) multivariate statistical process monitoring models for real-time process monitoring, and (iv) advanced process control. We discuss model development, implementation strategy and lessons learned, with particular emphasis on the end-to-end systems model. A demonstration of the modeling capability is provided for a multi-specific antibody.

The iSKID downstream process runs continuous protein A capture with periodic cycles of continuous low pH inactivation, anion exchange chromatography and single pass TTF into a collection vessel across several days from a perfusion bioreactor. Key learnings gathered from GMP manufacture of three different products include automation improvements for enhanced productivity, diversion control strategies, and process impurity clearance. These learnings help further optimize iSKID integration within the clinical manufacturing facility.

Intensified, continuous manufacturing processes have been developed to address challenges in rAAV production, improving scalability, yield, and efficiency. We present an upstream perfusion platform optimized for high cell density, continuous harvest, and feeding strategies, along with transient transfection optimization to enhance productivity. These innovations lay the foundation for overcoming current production limitations, ensuring consistent, high-quality rAAV for large-scale gene therapy manufacturing and advancing future therapeutic applications.

Perfusion and intensified cultures reduce costs, shorten production durations, and maximize recombinant productivity. Using NISTCHO, an open-access living standard, we achieved a 3x productivity increase with shorter culture durations through media screening, culture additives, optimized feeds, and seeding/bleeding strategies. Improved specific productivity via N-1 culture and feed strategies highlights their role in cell culture optimization for continuous and intensified processes.