The limited number of well-characterized model bacteria cannot address all the challenges in a circular bioeconomy. Therefore, there is a growing demand for novel platform organisms with enhanced resistance to extreme conditions, versatile metabolic capabilities, and the ability to utilize cost-effective renewable resources while efficiently generating desirable biobased products. Gram-negative (G-) thermophilic bacteria are attractive microbial platforms, promising significant advances in industrial and environmental applications and bioprocess robustness. 
 
This lecture will summarize our recent effort in developing and applying genetic and systems metabolic engineering tools for Caldimonas thermodepolymerans, a facultative G- thermophile that efficiently converts a spectrum of plant biomass sugars into high quantities of polyhydroxyalkanoates—fully biodegradable substitutes for synthetic plastics—and other valuable bioproducts. Due to its thermophilic nature, polymer production capacity, and environmental resilience, this bacterium is an attractive candidate for next-generation industrial bioprocesses.
 
Our current work includes optimizing the DNA transformation protocol, developing and testing plasmid-based tools for genome editing, including thermophilic CRISPR/Cas technology, developing a genome-scale metabolic model, and making initial progress in engineering chassis strains with enhanced bioproduction properties and facilitated genetic manipulation. Our work pioneers the biotechnological domestication of C. thermodepolymerans and related G- thermophiles, paving the way for better understanding their extremophilic nature and boosting their bioproduction potential.

Learning Objectives:

1.  Discover how thermophiles can contribute to bioprocess robustness and sustainability.

2. Discuss what the major bottlenecks are when domesticating non-canonical microbial hosts.

3. Evaluate the properties of bacterial bioplastics: polyhydroxyalkanoates.

Bioprocessing remains one of the key bottlenecks in translating advanced biotherapeutics into clinical reality. Whether developing biologics, viral vectors, or ATMPs, early-stage CMC activities are often dominated by trial-and-error experimentation driven by limited prior experience and fragmented datasets. The result is extended timelines, high experimental burden, and increased development risk.

While Quality by Design aims to enable structured and knowledge-based development, its implementation at early stages is frequently constrained by insufficient data and process understanding. This raises a central question: how can bioprocessing become more predictive and less empirical?

This presentation introduces a knowledge-driven workflow that integrates a structured process database with targeted experiments and modeling. Instead of broad trial-and-error studies, experience and data guide focused, high-value experimentation. Experimental results are iteratively combined with hybrid models to simulate process behavior and define only the most informative next experiments.

Using a viral vector case study, the session illustrates how this approach reduces unnecessary laboratory work, accelerates learning cycles, and improves decision-making. While demonstrated in viral vector development, the principles are broadly applicable across biologics, advanced therapies, and bioprocess-based validation or scale-up activities.

Participants will gain practical insight into how modern bioprocessing can shift from reactive experimentation toward structured, data-informed strategy.

Learning Objectives:

1. Identify major bottlenecks in early-stage bioprocessing and CMC.

2. Explain limitations of early QbD implementation when data is limited.

3. Describe how structured knowledge bases and modeling reduce experimental burden and improve decisions.

Join Dr. Mikael Kubista, a pioneer of developing and standardizing real-time quantitative PCR (qPCR) and digital PCR (dPCR) assays for research and diagnostics. As a member of the American Association of Pharmaceutical Scientists (AAPS) working group on PCR assay standards, Dr. Kubista will go into an in-depth discussion on best practices in PCR assay design, validation, and application within the context of cell and gene therapies. 

This session will provide practical, standards-based guidance informed by Dr. Kubista’s extensive experience and involvement in shaping industry recommendations. Attendees will gain insights into designing and validating PCR assays for critical applications such as biodistribution, transgene expression, viral shedding, and cellular kinetics. The webinar will also explore how to optimize assay performance, improve measurement accuracy, and determine when to use qPCR and dPCR technologies. In addition, Dr. Kubista will discuss emerging considerations for pharmacokinetic assays of LNP-mRNA drug products using RT-qPCR, an area of growing importance in the field. 

Whether you're involved in assay development, regulatory submissions, or therapeutic research, this webinar will equip you with actionable knowledge to align your work with current guidelines and accelerate progress in cell and gene therapy development. 

Learning Objectives:

1. Best practices in qPCR and dPCR assay design, validation, and application

2. How to design and validate PCR assays specifically for cell and gene therapy workflows

3. Tips for optimizing assay performance and selecting between qPCR and dPCR

Join this year's poster presenters in the Poster Hall during the Poster Discussion, Wednesday, April 1st, from 12:00pm – 1:00pm PST, to chat live about their posters and latest research and findings.

Lipid-laden macrophages are a defining pathological feature across a broad range of inflammatory diseases, including atherosclerosis, metabolic disorders, and chronic tissue injury, yet effective strategies to selectively modulate these cells remain limited. We recently reported an LDL-binding anti-inflammatory cytokine fusion protein that leverages endogenous lipoprotein trafficking to target inflammatory macrophages and suppress pathological activation. Building on this work, we are now developing a modular framework to systematically tune key biophysical and pharmacological properties of LDL-binding biotherapeutics. Specifically, we alter binding avidity by controlling the number of LDL-binding Fab domains, modulate signaling stoichiometry by varying the number of IL-10 payloads per construct, and extend circulating half-life through fusion to serum albumin. In parallel, we are establishing three-dimensional in vitro models of foam cell formation using gelatin methacrylate hydrogels that recapitulate lipid accumulation and macrophage phenotypic heterogeneity in a non-animal setting. Together, this integrated approach links precise physical characterization of LDL-binding biotherapeutics with functional evaluation in physiologically relevant 3D macrophage models, enabling early efficacy testing and rational optimization of immunomodulatory protein therapeutics.

Learning Objectives:

1. Describe how lipid-laden macrophages arise in inflammatory diseases.

2. Evaluate whether LDL-binding is an effective strategy for selectively targeting these cells.

3. Explain how biophysical design parameters of LDL-binding therapeutics affect function.

As demand for recombinant therapeutic proteins increases, there is continued emphasis on shortening timelines for cell line selection and increasing productivity by cell line engineering. Classical approaches, including random integration and antibiotic selection, result in a large number of low productivity clones and an extended cell line development time. Identifying high productivity clones necessitates screening a large number of clones, i.e. searching for a needle in a haystack. Antibiotic selection is time consuming and does not necessarily result in the highest clone productivity possible. In this presentation, I will discuss approaches for cell line selection including high throughput screening technologies, targeted transgene integration and a novel cell line selection approach based on transcriptional selection. 

Learning Objectives:

1. Explain the limitations of traditional random integration and antibiotic-based selection methods in biotherapeutic cell line development.

2. Compare high-throughput screening, targeted transgene integration, and transcriptional selection approaches for identifying high-producing clones.

3. Evaluate how modern host and clone engineering strategies can shorten timelines and improve productivity in recombinant protein manufacturing.

AlphaFold was published in 2021 and seemingly overnight effectively solved the protein folding problem, which had been an open challenge for decades. Since then, AI and ML methods have transformed the fields of (computational) protein science and engineering, culminating in the Nobel Prize being awarded in 2024 to the inventors of AlphaFold and the pioneer in this area. Thanks to these developments, it is now possible to start answering meaningful engineering questions about proteins, like what features should a protein have to best accomplish a particular task?

The PROTEIN PANT(z) Lab has been investigating interfaces between proteins to identify what features are important to protein binding with an eye towards enabling the rational design of binding proteins. Through investigating the stabilization of pairs of interacting residues in protein interfaces, we have identified novel features that appear to be critical to protein binding. This talk will discuss the development of those features and their applications in an end-to-end binding protein design pipeline that includes automated epitope identification, initial structure prediction, computational affinity maturation, and prediction of experimental viability.

Learning Objectives:

1. Define different features of residue-residue interactions at protein interfaces

2. Explain the importance of considering features of residue-residue interactions rather than just their existance

3. Describe applications of using features of residue-residue interactions in protein design

The session will explore the market growth of complex molecules, driven by advances in personalized medicine and biotechnology. We will identify key drivers and discuss the primary challenges in purification, including purity levels and process efficiency.

The presentation will also offer insights into innovative strategies and technologies for overcoming these hurdles and ensuring high-quality biopharmaceutical products. Join us to gain valuable knowledge and tools for navigating the complexities of purifying advanced biopharmaceutical products.

Learning Objectives:

1. Learners will understand how the rapid growth of complex and mAb‑based molecules is increasing purification challenges, and why yield, stability, impurity clearance, and scalability are now critical considerations in bioprocessing.

2. Learners will be able to explain how mild, higher‑pH elution strategies can improve purification outcomes, including higher yields, improved long‑term stability, and enhanced impurity and viral clearance.

3. Learners will recognize the importance of combining robust purification technologies with expert application support to enable reliable process scale‑up from development through commercial manufacturing.

In vivo CAR-T therapies represent a significant shift from traditional ex-vivo cell manufacturing by enabling direct genetic reprogramming of immune cells within the patient. While this approach improves scalability and accessibility, it introduces unique analytical and control strategies focused on vector quality, delivery efficiency, biodistribution, and immune activation.

This presentation will outline a risk-based analytical framework for in vivo CAR-T development, including identification of critical quality attributes, vector characterization, potency assessment, and in vivo pharmacodynamic monitoring. Emphasis will be placed on phase -appropriate testing strategies aligned with FDA lifecycle expectations for cell and gene therapies, enabling robust product understanding while supporting clinical development and regulatory compliance.

Learning Objectives:

1. Identify key critical quality attributes (CQAs) unique to in vivo CAR-T products compared to traditional ex vivo CAR-T

2. Design a phase-appropriate, risk-based analytical control framework integrating vector characterization, potency assessment and in-vivo monitoring tools

3.  Analytical refinement for in vivo CAR-T programs through the regulatory landscape

One obstacle associated with CAR T cells is their limited controllability after administration to the patient, which becomes particularly problematic in the case of severe toxicities. To be able to control CAR T cell function in vivo, we generated caffeine-responsive molecular switches by using high-end protein engineering. We demonstrate that these switches are clean (i.e. no background activation in the absence of caffeine) and efficiently turned on upon addition of caffeine. Furthermore, integration of these switch modules into CARs yields switchable “CaffCARs”, whose function can be precisely controlled via administration of caffeine. We show that this switch platform is modular, enabling caffeine-dependent regulation of CARs directed against different antigens (including antigens for leukemia, lymphoma and solid tumors). Importantly, these CaffCAR T cells are efficiently activated in response to caffeine concentrations that are achieved in human serum after drinking one cup of coffee. Finally, when we tested our CaffCAR T cells in a lymphoma mouse model in vivo, we observed high anti-tumor potency that could be regulated efficiently via administration of caffeine to the mice. Together, we introduce caffeine-responsive switch modules that enable functional regulation of CAR T cells in vivo with caffeine concentrations that are achieved in human serum after drinking one cup of coffee. 

Learning Objectives:

1. Define why we need functional control of CAR T cells? 

2. Describe the engineering strategy of drug-responsive molecular switches

3. Discuss optimal strategies to incorporate such switches into CARs

Structure is inherently important to the function and performance of many things in the world we live in. This extends to the materials used to manufacture and purify essential medicines needed to save and improve lives. This talk focuses on how a better understanding of structural properties in this engineering space can be applied to better design these complex, multi-stage devices. The goal of this body of work is to improve efficiency, effectiveness and sustainability in order to drive down costs, making modern therapies that pass through the clinic to the market accessible and affordable to all that need them.

The research focus will be examining the role of multiple imaging technologies, and how they can be applied in a complementary manner to better comprehend how purification material geometries can be evaluated and customized towards the specific chemical and physical needs of complex biological products. We will explore imaging techniques including confocal microscopy and X-ray computed tomography, and newer purification materials such as those made through nanofibers and 3D printing. More customizable options, in combination with a greater comprehension of how structure influences performance opens possibilities to improve medicines manufacturing of the past, present and future.

Learning Objectives:

1. Discuss the relationship between structure with function and performance

2. Recognize the role of purification materials and their associated properties

3. Identify potential uses for new materials in the medicines manufacturing space

Cell-based therapies have the potential to provide sustained, physiologically regulated delivery of therapeutic proteins, but their clinical translation has been limited by immune rejection, fibrosis, hypoxia, and scale-dependent transport constraints. Immunoprotective encapsulation technologies offer a promising strategy to overcome these barriers while avoiding systemic immunosuppression.
This presentation will provide an overview of micro- and macroencapsulation approaches for cell-based therapies, highlighting advances in biomaterial design, device engineering, and translational evaluation. Recent progress in chemically modified alginate systems designed to mitigate foreign body responses will be discussed alongside preclinical findings examining immune responses to xenogeneic and allogeneic grafts in large animal models. The talk will also explore macroencapsulation device strategies, including oxygenation approaches and implantation site considerations, to address mass transport limitations and maintain graft viability at clinically relevant scales.
By integrating biomaterials science with device engineering and large-animal translation, these advances aim to enable durable, immune-protected cellular therapies for diseases such as diabetes and other conditions requiring sustained protein delivery.

Learning Objectives:

1. Explain key barriers limiting translation of cell encapsulation therapies.

2. Discuss the advantages and disadvantages of both micro- and macroencapsulation strategies for cell therapies.

3. Identify 3 strategies currently being explored to improve effectiveness of encapsulated cell therapies.

Our society urgently requires a transition from fossil fuel dependence to a biosustainable economy. Engineering assembly-line or multi-domain megasynthases, such as type I polyketide synthases (PKSs), holds significant promise for replacing parts of petroleum-based chemical synthesis. We have developed a modular PKS-based platform that integrates a versatile loading module and terminal thioester reductases, enabling the production of diverse medium- and branched-chain diols, amino alcohols, and hydroxy acids with programmable access to branched structures at high titers in Streptomyces. Additionally, we further extended PKS applications to the sustainable synthesis of high-energy polycyclopropanated fatty acids (POP-FAs) as next-generation biofuels. Through engineering of an iterative PKS (iPKS) system, informed by computational modeling and domain swapping, we achieved the microbial production of designer POP-FAs with precise control over chain length and cyclopropanation, and with superior energy density and freezing point characteristics for sustainable aviation fuels. Collectively, these studies demonstrate the transformative potential of PKS engineering to access new chemical space and enable the biosynthesis of tailor-made molecules, from specialty chemicals to advanced biofuels.

Learning Objectives:

1. Explain the principles of polyketide synthase (PKS) engineering

2. Describe how PKSs can be engineered to produce tailor-made diols and high-energy biofuels

3. Assess the potential of PKS-based biosynthesis for replacing petroleum-derived synthesis