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Manuel J.T. Carrondo, PhD
Instituto de Biologia Experimental e Tecnológica (iBET)
Clifton E. McPherson, PhD
Protein Sciences Corporation, A Sanofi Company
Otto-Wilhelm Merten, PhD
Amine A. Kamen, PhD
McGill University
Martin A. Giedlin, PhD
Novartis Pharmaceuticals Corporation
Gary K. Lee, PhD
Sangamo BioSciences, Inc.

Dominic Esposito, PhD
Director, Protein Expression Laboratory
Frederick National Laboratory for Cancer Research
The Complete Genome Sequence of a Trichoplusia ni Cell Line, Tni-FNL


We have determined the complete genome sequence of a Trichoplusia ni cell line, Tni-FNL, to the highest quality and completeness of any lepidopteran organism sequenced to date. We will present data on the methodology used to map the genomic sequence along with bioinformatic analysis discussing similarities and differences among the sequenced lepidopteran hosts. In addition, we will discuss some of the unique characteristics of this cell line which may be responsible for its ability to produce higher levels of recombinant proteins than other cell lines, including commercially available lines such as High Five.


Dr. Esposito is currently the Director of the Protein Expression Laboratory (PEL) at the Frederick National Laboratory for Cancer Research (FNL) in Maryland. The PEL primarily generates DNA and protein reagents for the NCI national mission to target KRAS-driven cancers, as well as supporting NCI and NIH intramural investigators in production of difficult proteins. Prior to his role as director, Dr. Esposito led the Clone Optimization Group in the PEL for nine years and was responsible for the generation of over 15,000 expression clones, 400 new expression vectors, and several technological innovations in protein expression. Dr. Esposito received his BA in Chemistry at La Salle University in Philadelphia, and his PhD in Biochemistry at the Johns Hopkins University Bloomberg School. Dr. Esposito previously worked for Life Technologies, where he helped to develop the Gateway recombinational cloning system.

Lawrence C. Thompson, PhD
Principal Scientist and Group Leader, BioTherapeutics Pharmaceutical Sciences,
Analytical Research and Development, Pfizer, Inc.
Characterization of Viral Vectors


Viral vectors are extremely complex biological machines, making preparation of robust characterization packages a daunting task. Strategies and the associated analytics can be grouped into two categories: those that probe the nature of the intact particle and those that dig deeper into the molecular attributes of its components. Both are value added and when combined can be used to explore lot-to-lot comparability and explain the “why” behind movement in stability indicating assays.


Lawrence (Larry) C. Thompson, PhD is a Principal Scientist in Analytical Research and Development within BioTherapeutic Pharmaceutical Sciences at Pfizer. He has been with Pfizer for four years and is currently analytical lead on viral and plasmid-based immunotherapeutics. Previously, he spent three years in small biotech at two different companies as the analytical lead in the development of serum-based cancer diagnostics. He is a graduate of the University of Tennessee (Chattanooga) with a BS in Chemistry in 2001, and Vanderbilt University with a PhD in Biochemistry in 2006. He did his postdoctoral work at the University of Tennessee (Knoxville) from 2006–2010. His education and research path covers a wide breadth including small molecule synthesis; protein cloning, expression and purification; enzyme structure/function determination; protein-protein interaction investigation; serum antigen identification and mAb production/selection; and virus/plasmid method development and characterization (to name a few), generating a number of peer reviewed publications and presentations at scientific conferences as well as internally within Pfizer.

António M. Roldao, PhD
Senior Scientist, Instituto de Biologia Experimental e Tecnológica (iBET)
Bioprocess Engineering of Insect Cells for Pseudotyped VLP Expression and Optimization


Conformational-complex membrane proteins (MPs) are vaccine/drug targets in many diseases, but drug and vaccine development has been slowed down by the lack of efficient production tools. Co-expression of MPs with matrix proteins from enveloped viruses is a promising approach to obtain correctly folded proteins at the surface of ordered nanoscale architectures such as virus-like particles (VLPs), preserving their native lipidic environment.

In this work, we implemented an innovative site-specific recombination strategy based on flipase-mediated cassette exchange technology to establish reusable insect cell platforms for fast production of enveloped VLPs pseudo-typed with target MPs. Influenza M1 and HIV Gag proteins were evaluated as scaffolds, and proof-of-concept (PoC) demonstrated using two membrane proteins, the influenza HA protein (e.g. for vaccines) and the human beta-2 adrenergic receptor (e.g. for drug screening or antibody discovery). Bioprocess engineering schemes were designed (adaptive laboratory evolution to hypothermic culture conditions and supplementation with productivity enhancers), allowing improvement of HIV Gag-VLP production in the developed stable insect cells. Under hypothermic culture conditions, adapted cells expressed up to 30-fold more HIV Gag-VLPs than non-adapted cells. Noteworthy, the element driving such increase in productivity is the adaptation process and not the temperature shift as the latter alone leads to lower production yields. A more modest increase in productivity (up to seven-fold) was observed when supplementing non-adapted cell cultures with productivity enhancers NaBu and DMSO. PoC was successfully demonstrated in 0.5 L stirred-tank bioreactors.

Profiting from the platforms developed above, a modular system comprising stable and baculovirus-mediated expression in insect cells was established for the production of a multi-HA influenza VLP as vaccine candidate that otherwise could not be obtained due to baculovirus vector instability. By combining stable with transient expression systems, we could rationally distribute the number of genes to be expressed per platform and thus generate the target VLP for subsequent animal studies. In addition, a tailor-made refeed strategy was designed based on the exhaustion of key nutrients during cell growth resulting in a four-fold increase in HA titers per mL. PoC was successfully demonstrated in 2 L stirred-tank bioreactors. Overall, the insect cell platforms and bioprocess engineering strategies herein assembled have the potential to assist/accelerate drug and vaccine development.


António Roldao is a Chemical Engineer with a PhD in Engineering and Technology Sciences - Systems Biology (2010) from ITQB-UNL (Portugal). From 2010 to 2014 he was a senior researcher and director of fermentation technology at SysBio group headed by Dr. Jens Nielsen at Chalmers University of Technology (Sweden). In 2014 he became a senior researcher at the Animal Cell Technology Unit headed by Prof. Paula Alves and Prof. Manuel Carrondo at iBET / ITQB-NOVA (Portugal), and since 2015 he has been Investigador FCT. António has been awarded with several research grants, is an author of 18 scientific manuscripts in peer-reviewed journals, author or co-author of seven book chapters/conference proceedings, and has given over 25 oral and poster communications. António has been involved in many Portuguese and EU funded research projects. Current research focuses on the development of novel complex biologics with an impact in Human Health, e.g. VLP-based vaccines against infectious diseases such as influenza and dengue. To accomplish such objectives, bioprocess engineering and bottom-up systems biology approaches are combined with process monitoring and product characterization, thus undoubtedly fastening the generation of such products.

Steven E. Pincus, PhD
Associate Vice President Virology and Analytical Methods Development
FUJIFILM Diosynth Biotechnologies Texas, LLC
A Suspension Vero Cell Line for Production of Viral Vaccines and Viral Therapeutics


As the acceptance of viral vectors as a delivery system for therapeutics grows, biomanufacturers are looking for an alternative to the classical adherent cell production models. Adherent cell lines such as Vero and Madin Darby canine kidney (MDCK) are licensed for vaccine production, but they require large volumes of media, specialized large-scale adherent culture vessels with a large working footprint, and the equipment to support them. Suspension cell lines, such as Chinese hamster ovary (CHO) and HEK-293 are licensed for antibody production, but have not yet been licensed for vaccine production. A mammalian cell line that can simultaneously grow in suspension, support many types of viruses and viral vectors, and produce high virus titers would be of great value for production of vaccines and viral therapeutics.

Fujifilm Diosynth Biotechnologies Texas (FDBT) is a contract development and manufacturing organization that specializes in the production of viral therapeutics and vaccines. We have employed the use of a Vero cell line, which is permissive to a wide array of viruses, to create a platform for virus and viral vector growth. We have adapted this adherent Vero cell line to a serum-free, fast growing suspension culture. We have evaluated the ability of the adherent and suspension cell lines to amplify viruses and viral vectors such as influenza virus, adeno-associated virus, and adenovirus. This proprietary suspension Vero line will be an excellent asset to viral vector and vaccine manufacturing, as it will allow the growth of commonly difficult-to-scale vectors. FDBT has also developed a wide variety of analytical methods to evaluate the growth of virus and viral vectors which are commonly used to support the numerous manufacturing stages of vaccine production. These include analyses designed to identify (PCR, ELISA), quantify (FACS, immunostaining, plaque assay), and verify the purity (HPLC, SDS-PAGE/Western, ELISA) of our products.


Steven Pincus is Associate Vice President of Virology and Analytical Development at Fujifilm Diosynth Biotechnologies Texas where he leads teams involved in process development and analytical development for client driven programs in viral therapeutics and vaccines. He obtained his BS and PhD in Biochemistry from the State University of New York at Buffalo. His PhD thesis project involved studies on the mechanism of adenovirus DNA replication, and he gained experience growing, purifying, and titering multiple adenovirus serotypes. He obtained his postdoctoral training in the microbiology department at the State University of New York at Stony Brook under an NIH fellowship studying inhibitors of picornavirus replication, and gained experience in the growth, purification, and titration of poliovirus and the growth of other picornaviruses.

He then joined Virogenetics where he was Senior Scientist and Platform Leader in Molecular Biology and Microbiology. At Virogenetics he was involved in the development of highly attenuated poxvirus vaccine vectors (NYVAC, ALVAC, TROVAC) and developed vaccine candidates for measles virus, flaviviruses (JEV, dengue, yellow fever), human cytomegalovirus, and therapeutic vaccines against several cancers along with development of molecular assays necessary for releasing clinical lots. At Elusys Therapeutics he was Senior Director Virology and Animal Biology and received $1.6 million NIH grant funding to investigate the use of the Elusys antibody heteroplymer technology as a therapeutic for vaccinia vaccination complications. He developed animal models and release assays for heteropolymer clinical trials and supported the development of an anthrax anti-toxin monoclonal that is funded by the Biomedical Advanced Research and Development Authority (BARDA). He then joined Novavax, Inc., a biotech focused on the development of vaccines against influenza and respiratory syncytial virus (RSV), based on virus-like particle (VLP) platform vaccine technology. There he was responsible for analytical development, quality control and quality assurance, and recombinant baculovirus production. He was a key member of the team that secured a contract award valued at up to $179 million by BARDA for the advanced clinical and manufacturing development of recombinant vaccines for the prevention of seasonal and pandemic influenza. He has consulted for several companies in the areas of assay development, transfer, qualification, and validation for monoclonal antibody and vaccine projects.

Christoph Geisler, PhD
Chief Research Scientist, GlycoBac LLC
Adventitious Viruses Contaminating Insect Cell Lines


Starting in the late 1970s, a wide variety of viruses had been found to contaminate insect cell lines. Surprisingly, adventitious viruses contaminating insect cell lines routinely used for recombinant protein and AAV production were not discovered until 2007 (High FiveTM) and 2014 (Sf9). These discoveries raised important questions regarding the biosafety of therapeutics produced in this platform. In this talk, I will discuss viruses that can contaminate insect cell lines, including several relatively little-known ones. As most persistent viral infections of insect cell lines were discovered serendipitously, I will also discuss how these viruses were discovered as well as detection methods that can be used to probe for viral contamination. Finally, I will present new virus-free insect cell lines that have been developed as alternatives to contaminated cell lines, and to what extent these can serve as viable alternatives to their well-established, contaminated counterparts.


Christoph Geisler has worked in the baculovirus insect cell system since 2005. He has experience in both cell line development and baculoviral vector engineering, and specializes in the glycobiology of the system. Recently, he has developed new bioinformatics approaches to screen for adventitious viral contaminants in cell lines used to produce biopharmaceuticals.

Francesca Bellintani
Downstream Vector Development Manager, MolMed S.p.A.
Retroviral and Lentiviral Production in Disposable Bioreactor: Development
and Scaling Up of Upstream and Downstream Process Steps


MolMed is a medical biotechnology company focused on research, development, and clinical validation of innovative therapies to treat cancer and rare genetic diseases. MolMed's portfolio in cell and gene therapy includes anti-tumor drugs in clinical and preclinical development:
- Zalmoxis® (TK) is a cell-based therapy enabling bone marrow transplants from partially compatible donors, in absence of post-transplant immune-suppression, currently in Phase III in high-risk acute leukaemia and approved by European Medicines Agency (EMA) for conditional marketing authorization;
- CAR-CD44v6, an immuno-gene therapy project potentially effective for many haematological malignancies and several epithelial tumours, currently in preclinical development.

Moreover, MolMed offers high level expertise to develop, perform, and validate custom studies, optimize and scale-up manufacturing projects, devise innovative testing procedures, and address the unique test specifications required for novel therapies. In order to meet vectors manufacturing demands for both clinical and commercial phases, MolMed is developing modular innovative processes (24 L, 48 L, 200 L) in cell factories and in disposable bioreactors that permit an increase in lentiviral/retroviral (LV/RV) vector productivity, reducing cost of goods and preserving high vectors quality. MolMed is also investigating LV production using suspension cell lines cultured in the absence of animal-derived components to ensure a higher level of safety and unlimited possibilities for scale-up. MolMed is improving hematopoietic stem cell and T-cell production processes in order to develop closed systems with a high level of cell transduction, lower number of manipulations to increase sterility assurance levels, and process reproducibility.


Francesca Bellintani is Downstream Process Development Manager at MolMed. She holds a degree in Biology from the University of Milan for her work on overproduction and purification of recombinant proteins. She specialised in biochemistry working on phage display while at Cambridge University. She joined MolMed in 2007 and has worked in the Development Unit with growing responsibilities. She has strong operations expertise in the development of processes for the purification of proteins and retroviral/lentiviral vectors using chromatography and tangential flow filtration techniques. Francesca supervises a multidisciplinary team of scientists on projects from development to early phase GMP manufacturing. Her knowledge includes ICH guidelines for development and qualification of analytical methods to assess identity, potency, impurity profile, and stability (UPLC, SDS-PAGE, Western blot, and ELISA).

Kendra Steele, PhD
Senior Research Scientist, ParaTechs Corporation
Simplifying Membrane Protein Purification: Introducing a Fluorescent BEVS Protein System that Enhances Protein Production and Greatly Simplifies Detergent Screening


Membrane proteins are central and essential to cellular structure, metabolism, and function. Consequently, membrane protein mutations frequently cause disease and are often therapeutic targets. Over 60% of today’s drug targets are membrane proteins with 30% of those treatments targeting G-protein coupled receptors (GPCRs), an important family of cell surface proteins. Despite enormous growth potential and advances in crystallography, mass spectrometry, and protein-ligand interactions, GPCR therapeutics remain difficult to develop, largely because GPCRs are difficult to isolate in intact and functional forms. Purification methods are often cumbersome, time-consuming, and ineffectual, hampering GPCR purification and downstream experimentation. Thus, rapid and simplified methods that can be tailored for purification of specific membrane proteins would greatly enhance opportunities in this important therapeutic sector.

The challenge to purifying membrane proteins is two-fold: to produce large quantities of pure, properly processed protein, and to solubilize active protein from membranes. Mammalian membrane proteins, and notably GPCRs, are often successfully produced with the baculovirus expression vector system (BEVS) in insect cells. ParaTechs has improved the BEVS by introducing viral ankyrin proteins (vankyrins) that delay apoptosis of baculovirus-infected insect cells, thereby enabling prolonged synthesis, accumulation, and processing of complex recombinant proteins. But the ability to solubilize and isolate the membrane protein in a native, functional form is perhaps even more important than achieving a high level of protein production. A major impediment to membrane protein research is simply finding the right detergent and conditions to purify your protein. Identifying the right detergent typically begins with an uninformed screen of detergents selected with inadequate knowledge of the proteins physio-chemical properties. Commercially available detergent panels do not systematically manage membrane protein purification, isolation, and analyses. Kit users are required to develop, process, and analyze each sample by methods unspecified by the kit. For example, if a user purchases a kit with 96 detergents, they must perform 11 Western blots to screen each sample (if using a 10-lane polyacrylamide gel with molecular weight marker). Here, we introduce VELucity, a fluorescent protein system that enhances production, improves stability, and enables screening for solubilization of proteins in a panel of commercial detergents before protein purification. This system will allow researchers to rapidly determine the right detergents for their protein by visual fluorescence, thereby reducing screening time from days to minutes.


Dr. Steele is the lead scientist on baculovirus protein expression projects at ParaTechs Corporation. She is the creator of VELucity, a fluorescent protein system that enhances membrane protein production, improves stability, and enables screening for solubilization of proteins in a panel of commercial detergents before protein purification. She also runs the contract program that expresses a user’s protein using the vankyrin enhanced-baculovirus expression system.

Nicole Faust, PhD
Chief Scientific Officer, Cevec Pharmaceuticals GmbH
CAP-GT, a Platform Addressing the Production Challenge


Nicole Faust is Chief Scientific Officer and Managing Director at CEVEC Pharmaceuticals, overseeing the company´s activities in gene therapy vector production and recombinant glycoprotein expression. Over the last 18 years she has held scientific management positions with several biotech companies. Before joining CEVEC in 2011, she worked with Lonza as Director R&D combining her expertise in cell biology and gene transfer technology to develop cell-based assays. Prior to Lonza she was heading the Molecular Technology Department at Artemis/Taconic Biosciences, using site-specific recombination and gene editing technologies for the development of pharmaceutically relevant animal models. Nicole holds an MBA degree from Educatis University, Switzerland and she received her PhD in Molecular and Cell Biology from University of Freiburg and spent her postdoctoral period at EMBL, Heidelberg, where she worked on embryonic stem cells and cell differentiation within the hematopoietic system.

Philip H. Coelho
Chief Technology Officer, Thermogenesis Corp.
How Cell Processing Systems Provide Automated
“Functionally Closed” CAR T-Cell Production


Philip H. Coelho, Founder and Chief Technology Officer (CTO) of Thermogenesis Corp., is an engineer/inventor, educated at the University of California, Davis, where he specialized in thermodynamics and machine design. He has been awarded more than 40 US patents related to cell selection, cell cryopreservation, cell washing, automated cryogenic freezing and storage of cells, and harvesting clotting proteins from blood.

As Founder, Chairman, and CEO of Thermogenesis Corp., Mr. Coelho developed the Thermoline ultra rapid blood plasma freezers and thawers, the CryoSeal automated system for preparing fibrin sealant from a surgical patient’s blood, the BioArchive robotic system for controlled-rate freezing and cryogenically archiving units of hematopoietic stem and progenitor cells (HSPCs) derived from placental/cord blood, and the AutoXpress System which automates harvesting of HSPCs from cord blood into a 25 mL dual-compartment freezing bag. The BioArchive and AutoXpress Systems have established the highest quality GMP standards in cord blood processing and banking worldwide.

In 2002, Mr. Coelho initiated an effort to obtain federal funding sufficient to allow qualified public cord blood banks to build a substantial inventory of ethnically diverse, clinical grade, cord blood stem cell units for all patients in need. Passed unanimously by Congress and signed by President Bush on December 19, 2005 the “Stem Cell Therapeutic and Research Act of 2005” authorized and began financing a national inventory of 150,000 units of cryopreserved, HLA typed cord blood stem cells and a website that allowed transplant physicians to choose optimal stem cell units for the treatment of blood cancers such as leukemia, myeloma, and lymphoma; inherited immunodeficiencies and marrow failure disorders; and inherited disorders or errors of metabolism. Mr. Coelho retired from Thermogenesis Corp in August 2007.

Two years later, in 2009, Mr. Coelho co-founded SynGen Inc., and began the development of programmable systems that improve the speed and efficiency of the isolation and capture of purified stem, progenitor, and immune cells residing within peripheral or placental/cord blood, bone marrow aspirate, and leukapheresis. In July 2017 SynGen merged with Thermogenesis Corp, now a subsidiary of Cesca Therapeutics, and Mr. Coelho accepted the position of CTO in the combined company. He is now leading the development of CAR-TXpress, an automated method of manufacturing therapeutic doses of CAR T-cells in a functionally closed cell processing system.

Ana Sofia Coroadinha, PhD
Head of Cell Line Development and Molecular Biotechnology Laboratory
Instituto de Biologia Experimental e Tecnológica (iBET)
Novel Stable Lentiviral Vector Producer Cells: Overcoming Viral Vector Cytotoxicity
Etienne Boutry
Director Bioprocess R&D Europe, Sanofi Pasteur SA
Challenges in Process Development and Industrialization of Live Virus Vaccine:
Dengue and Flavivirus Experience on a Vero Cell Line
Lesley Chan, PhD
Scientist II, Vector Process Development & Manufacturing, bluebird bio Inc.
The Development and Intensification of a
Lentiviral Vector Manufacturing Process Using Stable Cell Lines
Hanna P. Lesch, PhD
Research and Development Director, FinVector Vision Therapies OY
Large-Scale Production Work of Adenoviral, Lentiviral, and AAV Vectors
Otto-Wilhelm Merten, PhD
Head of Applied Vectorology and Innovation, Généthon
Production of AAV Vectors Using the Insect Cell/Baculovirus System
Scot Shepard
Director, Biologics Process Design, Takeda Pharmaceuticals U.S.A., Inc.
Commercial Scale Manufacture of Virus-Like Particles to Supply
Global Clinical Trials of Takeda’s Norovirus Vaccine
Charlotte Dyring, PhD
COO & Co-Founder, ExpreS2ion Biotechnologies
Nathalie A. Clément, PhD
Associate Director, PGTC Vector Core Laboratory, University of Florida
Christopher W. Kemp, PhD
President, Kempbio, Inc.
Helen Maunder, PhD
Principal Scientist, Oxford BioMedica plc
James J. Mulé, PhD
Executive Vice President Applied Science
H. Lee Moffitt Cancer Center & Research Institute
Christine Le Bec, PhD
Head of Analytical Development, R&D, Généthon
Sergei Zolotukhin, PhD
Professor, Division of Cellular & Molecular Therapy, University of Florida
John T. Elliott
Cell Systems Science Group Leader, National Institute of Standards & Technology
David Hodl
President, Cell Processing Business Unit, SynGen Inc.
Gary K. Lee, PhD
Associate Director, Genome Editing, Sangamo BioSciences, Inc.
Kimberly Lounds-Foster
Corporate Vice President, Global Supply, Celgene Corporation
Laura A. Palomares, ScD
Researcher and Professor, Universidad Nacional Autónoma de México
Scott Estes, PhD
Head of Upstream Process Development & Manufacturing, Codiak BioSciences
Marc G. Aucoin, PhD
Associate Professor, Center for Bioengineering and Biotechnology
University of Waterloo
Thomas Wesley Powers
Senior Scientist, BioTherapeutics Pharmaceutical Sciences,
Worldwide Research & Development, Pfizer, Inc.
Avery D. Posey Jr.
Associate Laboratory Director, University of Pennsylvania
Chia-Hsing Pi
PhD Student, University of Minnesota
Tam Soden, PhD
Senior Director, Analytical Department, Product Sciences, Kite Pharma, Inc.
Gabriel Parra, PhD
Principal Investigator, Division of Viral Products, FDA CBER
Robin Levis, PhD
Deputy Director, Division of Viral Products, FDA CBER
Arifa S. Khan, PhD
Supervisory Microbiologist, FDA CBER
Thomas Meins
Founder, CRELUX GmbH
Jacek Lubelski, PhD
Director Vector & Process Development, uniQure N.V.

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