DNA vaccines have emerged as a promising technology for developing COVID-19 vaccines due to their ability to be quickly designed and manufactured after obtaining the pathogen or cancer antigen genome sequence. The recent approval of ZyCoV-D, the first DNA vaccine for COVID-19 for emergency use has demonstrated this efficacy.

However, utilizing novel technology platforms and development paradigms may increase the risks which requires careful safety evaluation. As a result, it is critical to ensure the quality of the naked plasmid from its source in order to satisfy the stringent requirements of DNA vaccines and drugs.

To this end, GenScript ProBio has developed a plasmid DNA platform and a viral vector platform, and has accumulated extensive expertise in manufacturing plasmids for various applications, including raw materials, drug substances, and even drug products. From preclinical to clinical to commercial supply, GenScript ProBio is dedicated to supporting the success of more gene and cell therapies.

Vaccines have played a major role in protecting human health as one of the key means of controlling infectious diseases. More than 20 high-risk diseases can be currently prevented through vaccination. The importance of vaccines was highlighted during the outbreak of COVID-19, when developing a vaccine was considered one of the most important ways to end the pandemic. It only took one year for mRNA vaccine to go from obscurity to popularity worldwide. In the post-COVID-19 era, mRNA vaccines still have the potential for substantial growth, especially in the field of tumor immunotherapy. In this webinar, we will review the main types of vaccines and the pros and cons of each type. We will discuss the key challenges for developing mRNA vaccines and the solutions GenScript can offer. Considerations and optimization steps that can be implemented to improve vaccine performance.

Learning Objectives:

1. List typical vaccine types and mechanisms. 

2. Discuss mRNA vaccines development process.

3. Discuss key technical hurdles in mRNA vaccine development.

Many commercially promising proteins are difficult-to-express (DTE) in living cells and are lost before an industrial production process can be established. Cell-free protein expression (CFPE) is well suited to producing DTE proteins but has largely been limited to microgram yields.

Here we present a eukaryotic lysate, sold as ALiCE® by LenioBio GmbH, which produces protein yields of up to 3mg/ml within a 48 hour reaction time. Being a eukaryotic system, ALiCE® has incredible promise for protein engineering due to its capabilities in performing post-translational modifications including disulfide bonds and N-glycosylation.

Recent developments in the scaling of ALiCE® bioprocessing now permit the production of liters of lysate, presenting a new challenge in scaling of the CFPE reaction to match.

We present case studies of cytosolic and a microsomally expressed proteins produced by ALiCE®, and describe their subsequent purification and characterization. We further disclose world first liter scale eukaryotic CFPE reactions using commercially available bioreactors.

Learning Objectives:

1. Discuss the difficulties of producing certain protein classes.

2. Explain why cell-free protein expression technology is more appropriate for producing certain classes of proteins.

3. Discuss factors important in scaling of cell-free protein expression for commercial scale protein manufacturing.

This presentation will discuss some of the challenges related to developing antibody therapeutics and how GenScript's comprehensive portfolio of services and products can increase the efficiency of the development. GenScript offers a full range of solutions, from target identification and validation to protein expression and purification of antibodies. Their state-of-the-art technologies, including antibody library construction, high-throughput screening, and in vitro maturation, enable rapid identification and optimization of lead candidates. In addition, GenScript's antibody engineering platform allows for customized modifications to enhance antibody function, stability, and specificity. With a focus on quality and reliability, GenScript provides end-to-end solutions that accelerate the drug development process and increase the chances of successful clinical outcomes.

Learning Objectives:

1. Discuss challenges in developing antibody therapeutics.

2. Discuss GenScript products and services to support lead generation and optimization.

3. Discuss GenScript products and services supporting preclinical and clinical stages

Allogenic cell therapies require the constant supply of billions of cells generated by robust, and economically viable bioprocesses. Human induced pluripotent stem cells (iPSCs) are a unique source for the production of functional human cell types, fueling advanced regenerative therapies. Single-Use stirred tank bioreactors have the potential to become the new gold standard for the commercialization of allogeneic cell therapy production. We here show, how we instrumented stirred tank bioreactor technology to take iPSC cultivation to the next level.

In this talk, we want to provide a deeper understanding of hPSC bioprocessing and how single-use stirred-tank bioreactors can aid in achieving this goal. Innovations in impeller designs improve the cultivation of stem cells as aggregates or on microcarriers. By precisely monitoring and controlling critical process parameters, we will show how bioreactor processyields can be significantly increased.

Learning Objectives:

1. Define basic principles of stem cell bioprocessing in stirred systems.

2. Identify the importance of process monitoring.

3. Process optimization: Discover the different possibilities of parameter control.

During my research career I have worked to further understanding of how intracellular organisation changes in response to the needs and environment of the living cell, focusing on the regulation and function of the actin and myosin cytoskeleton. Since joining the University of Kent School of Biosciences with a BBSRC David Phillips Research Fellowship, my research group has used molecular-genetics, biochemical and live-cell imaging techniques to study the regulation, mechanism and function of dynamic cellular components in growing and dividing cells. Using unicellular yeast and bacterial model systems, we have gained insights into universal mechanisms and regulatory processes of cell growth and division. Over recent years I have applied this expertise in microbial cell biology to develop a number of recombinant technologies for use in biotechnology applications. These bacterial technologies, which include recombinant amino-terminal acetylation, microcompartment utilisation and more recently recombinant vesicle production, are a main focus of my group's current research activities.

Learning Objectives:

1. Define what the challenges are in the expression of recombinant proteins in a cell.

2. List the advantages of using bacteria to express recombinant proteins.

3. Discuss what the current limitations are of using bacteria to produce recombinant protein.

From gene cloning and initial clone selection through to final cell evaluation, the continual assessment of cell count and viability is important for determining the best growing and highest producing clones. This information can be used for monitoring proliferation rates, optimizing growth conditions and normalizing cell data for further studies. Traditional methods for measuring cell count and viability are often low throughput, time-consuming and lacking in linearity. Advanced flow cytometry provides a fast, accurate and reproducible solution for cell viability and density quantification, which is essential for efficient biologics development processes.

This webinar will present data on how the high throughput iQue® advanced flow cytometry platform can be effectively used for simultaneous and accurate evaluation of cell count and viability.

Learning Objectives:

1. Explain how to achieve reproducible, quantitative analysis of cell count and viability.

2. Discuss reducing sample preparation time with a simplified workflow.

3. Discover ways to relieve clone selection bottlenecks with rapid and simultaneous evaluation of IgG titer, specific productivity and cell health.

The field of gene therapy has enabled the treatment and potential cures for serious, monogenetic diseases.  This form of advanced therapy relies on the delivery of the “correct” copy of a gene to a patient’s cells, and often utilizes a viral vector to achieve this goal.  This presentation will give an overview of the history of gene therapy products as well as insight into the current state of the market and the size of the pipeline.  In addition, a broad overview of the cGMP manufacturing process for adeno associated virus (AAV)-based gene therapies will be covered, including unit operations for production, purification, and formulation.  Finally, other relevant topics such as targeted drug delivery, navigating the regulatory process, and the price point of gene therapy products will also be covered.  

Learning Objectives:

1. Define what gene therapy products are.

2. Describe different types of vectors for gene therapy.

3. Outline a general process for AAV manufacturing.

The application of modeling tools to better understand and control bioprocess development and manufacturing has become a hot topic within the last few years. However, what do we actually need to create digital bioprocess twins? Is it just about data and powerful modeling algorithms, or is there more to consider to accurately describe a process, transfer the knowledge and model structure from early-stage development until manufacturing?

Discussion points and provided solutions:

•    The overall goal of process modeling
•    Experimental design concepts
•    Different modeling approaches for different requirements
•    Solutions to speed up both up- and downstream development
•    Model transfer along with scales
•    Model usage for real-time monitoring and control

Learning Objectives:

1. Describe what a digital bioprocess twin is and what can it serve for.

2. Discuss how to setup a digital twin and what are important things to consider.

3. Explain why the model structure is of importance to achieve both fast learning and process control goals.

In the past decade, there has been an increasing emphasis on translational research focused on cell and gene therapy. This has led to significant scientific advances in understanding disease mechanisms and developing novel, effective, and safe therapeutic interventions. Translational research includes the discovery of promising targets and lead compounds, preclinical studies, and attaining high-quality, consistent, and effective clinical product development. GenScript offers a wide selection of services and products, enabling a smooth transition of therapeutic development from Target Identification to Clinical Development. More specifically GenScript has made significant advancements in cell engineering, cell isolation and activation, and other related areas. Join us to learn how GenScript’s comprehensive solutions for cell and gene therapy will enhance your research and clinical development workflow.

Learning Objectives:

1. Discuss tools used for research and manufacturing for every step of CGT.

2. Describe methodologies for viral and non-viral gene editing.

3. Describe methods for automation of the cell isolation process.

Imagine, all of your lab work, your experimental data, your sample information and lineage tracing, your protocols and inventory management tools, all living on one, cohesive platform. What if you never had to worry about finding that long-lost file from 15 years ago or even had reports readily available at your fingertips for when the FDA shows up at your door? Sounds great right!? With digital lab platforms, you’ll have all the tools necessary to remain organized and compliant in any bioprocessing facility. 

There are many reasons to switch over to a digital lab platform, but here are 4 key points to highlight:

1. Centrality - Having a centralized, cloud-based location for all data related to your sample inventory and project/experiment results, making collaboration and archived information recall easier within your lab space. You can even integrate with other software and hardware in the lab!

2. Compliancy - Remain FDA compliant by automating workflows (electronic batch records, protocols, equipment management), removing human elements (mistakes), viewing/downloading audit trail repositories, assigning permission-based roles, data storage and redundancy backups, electronic witness signatures, etc.

3. Security - How secure is your data actually? Does your current system offer the following: ISO-27001 certification, GXP compliance, 21CFR Part 11 compliance, multi-server architecture, high availability installations, encrypted networks, on-site security, dedicated network security team, third-party penetration tests, redundancy and backups, disaster recovery procedures, single sign-on, two-factor authentication, and secure credential storage?

4. Efficiency - Make your lives easier by saving time and money! No more wasted efforts searching for lost data or documents, misplacing samples, stressing over compliance regulations, organizing your research, performing inventory, and so on and so forth. This is the end goal!

Learning Objectives:

1. Identify benefits of cloud based storage and how to centralize your data management system.

2. Discuss how to deal with compliance and security matters in the laboratory.

3. Discuss efficiency gains by means of digitalization and standardization.

High-resolution, 3D imaging enabled depth filters, chromatography columns and sterile filters to be analysed across multiple length scales. Generating a geometry-based database enabled structure to be related to function and performance has allowed for chemical and physical characteristics required for optimal separation to be identified. This presentation examines a range of downstream processing examples and how the information has been used to design and 3D print new materials.

Learning Objectives:

1. Describe methodologies for X-ray CT imaging of bioprocess structures.

2. Discuss how chromatography structure relates to function.

3. Compare conventional DSP materials to 3D printed structures.

In search of the next generation chromatographic technique for antibody purification, our team has developed an affinity chromatography method using the unconventional nucleotide binding site (NBS). By generating resins that display ligands that bind to the NBS that is conserved on all immunoglobulins, we achieve purity over 99% with >99% column efficiency, supporting that NBS method provides a stable, reusable, and inexpensive alternative for purification of humanized and chimeric antibodies. 

Learning Objectives:

1. Describe a not-so-well-known binding site on antibody structure.

2. Describe a novel affinity purification engineering for antibodies.

3. Describe quality control measures for evaluating antibody purity.

Protein production is a multi-billion dollar industry, with applications in pharmaceuticals, industrial enzyme use (eg in laundry detergent), and biomaterials. Bacteria are receiving renewed interest as protein production hosts because of their fast growth and tractability, but traditional production systems in bacteria are intracellular. Secretion is gaining traction as a strategy for bacterial production, as it enables continuous fermentation and reduces purification costs. To engineer these systems to be competitive with production in other microbes, researchers need high-throughput screens to probe the secretion system and amount of secreted protein. A popular choice is a fluorescent protein, but those derived from Aequorea victoria form a covalent bond upon formation of the chromophore, making secretion via systems that unfold the protein impractical. Other assays are highly dependent on environment, rendering them incompatible with various media additives. We present the development of a suite of fluorescence-based assays that overcome these challenges, based on the de novo designed mini Fluorescence Activating Protein (mFAP). This protein is expressed well, is compatbile with secretion in an unfolded state, and only becomes fluorescent upon addition of an inexpensive small molecule. We show that the assay provides a linear range over three orders of magnitude, and maintains linearity in backgrounds ranging from minimal to rich media. We demonstrate the utility of these screens to characterize and engineer the Salmonella enterica Type III Secretion System (T3SS). We identified variants that secrete non-native proteins at high titer, and engineered an optimized, more robust secreting strain of Salmonella. We are using the engineered strain for the high-titer production of a variety of biochemically challenging heterologous proteins, such as growth factors, antibodies, and toxic antimicrobial peptides. 

Learning Objectives: 

1. Explain the advantage of secreting a bioproduct, in the context of a manufacturing process.

2. Define the term fitness landscape in the context of protein engineering.

3. List and briefly describe the four types of assays commonly used to assess secretion titer.

Immune cell therapies hold tremendous potential in the treatment of cancer. Among immune cells, natural killer (NK) cells have a powerful innate ability to recognize and eliminate tumor cells. However, in the microenvironment of tumors, especially solid cancers, NK cell numbers are low, and those cells that are able to infiltrate these tumors are dysfunctional. In order to develop NK cell immunotherapies able to overcome aggressive mechanisms of suppression in tumors and enable them to more specifically target various cancers, genetic engineering approaches have been utilized to immunometabolically reprogram and enhance the function and specificity of NK cells against cancer. These genetically-engineered cell-based therapies have been formulated into clinical products and used to treat diverse cancers. This talk will give an overview of emerging engineered cell-based therapies, and how targeting mechanisms of immunometabolic suppression can yield functionally competent immune cells. Finally, the seminar will discuss how gene and cell engineering can be leveraged to generate powerful engineered NK cells, including multispecific and tumor-responsive variants, as well as new developments in the generation of allogeneic, off-the-shelf immunotherapies based on iPSC-derived NK cells, able to tackle complex mechanisms of treatment resistance in cancer. 

Learning Objectives:

1. Define the principles of cell-based therapies.

2. Outline approaches and targets for genetic engineering of immune cell therapies.

3. Describe applications and challenges of cell-based therapies in therapeutic settings.

Self-removing tags can be used for simple and effective purification of recombinant proteins and peptides. The key component of these tags is an intein, which is a protein element with the ability to excise itself out of a host protein. Engineered inteins have many applications, including the ability to detach themselves (self-cleave) from a fused target proteins in response to a small buffer pH change. This capability has significant applications in the field of protein purification, where one segment of a split intein can serve as a self-removing affinity tag. In this presentation, a novel self-removing tag will be discussed and compared to alternate methods. Case studies will also be presented, along with a deep dive into optimizing this type of protein purification process.

Learning Objectives:

1. Define what how self-removing tags are used during protein purification processes.

2. Describe protein purification steps.

3. Outline examples of proteins which were studied and their expression hosts.