COURSE DIRECTORS

Martin L. Yarmush, M.D., Ph.D., Professor, Department of Biomedical Engineering, Rutgers University; Director, Center for Engineering in Medicine, Massachusetts General Hospital

Martin Yarmush received B.S. degrees in Biology and Chemistry from Yeshiva University, Ph.D. degrees in Biochemistry and Chemical Engineering from Rockefeller University and MIT, respectively, and an MD degree from Yale University. Dr. Yarmush has served on the faculties of MIT, Rutgers and Harvard leading research programs in the areas of Molecular and Cellular Bioengineering, Tissue Engineering, and Metabolic Engineering. He is currently a Professor in the Department of Biomedical Engineering at Rutgers and also leads the Center for Engineering in Medicine at the Massachusetts General Hospital. Dr. Yarmush is well known at Rutgers as the founding Director of the Biotechnology Training Program.

Gregory Russotti, Ph.D., Director, Cellular Process Development, Celgene Cellular Therapeutics

Gregory Russotti is currently the Director of Cellular Process Development at Celgene Cellular Therapeutics in Warren, NJ. His group is responsible for developing, optimizing, and scaling up cell isolation, expansion, and formulation processes and transferring these technologies to clinical and commercial GMP manufacturing facilities. Celgene Cellular Therapeutics isolates their cells from human placentas after full-term healthy births and plans to use these cells to treat a variety of unmet medical needs. Prior to joining Celgene in 2006, Greg spent nearly 15 years at Merck Research Laboratories developing products that included live virus vaccines, monoclonal antibodies, recombinant vaccines, and microbially-produced natural products. He worked on development and scale-up of cell culture, microbial fermentation, and downstream isolation processes and was responsible, at one point, for transferring processes into a pilot plant and, in other roles, for transferring processes to manufacturing groups for clinical and commercial production, which included start-up of a new factory. Greg received his B.S. and M.S. degrees in Chemical Engineering from Rensselaer Polytechnic Institute. For his M.S. thesis he worked with Professor Georges Belfort on the design and construction of a novel perfusion mammalian cell bioreactor. Greg received his PhD in Chemical and Biochemical Engineering at Rutgers University while being supported by Merck’s Doctoral Fellowship program. During his PhD studies, under the guidance of Professor Martin Yarmush, he investigated the concept of inducing cold tolerance in mammalian systems through prior stress conditioning, the ultimate goal of which was to design improvements in the ways we preserve cells, tissues, and organs.

Bruno F. Marques, Ph.D., Senior Research Chemical Engineer, BioPurification Development, Bioprocess R&D, Merck Research Laboratories

Bruno Marques earned a B.S. in Chemical Engineering (with Specialization in Energy/Environment/Economics) from the Illinois Institute of Technology, and a Ph.D. in Chemical Engineering from Carnegie Mellon University. During graduate studies, under the guidance of Prof. James Schneider, he developed di-alkyl peptide nucleic acid amphiphiles that can be incorporated into surfactant microstructures, such as liposomes and micelles, and serve as oligonucleotide sequence tags in highly sensitive analytical devices. In 2005, he accepted a position with Merck's BioProcess R&D group, where he has been working on downstream processing of therapeutic proteins, with an emphasis on development and scale-up of monoclonal antibody (MAb) purification processes. In this role, he has led tech transfer efforts from the bench-scale to Merck's pilot plants, for GMP production of MAb drug product for clinical trials. More recently, Bruno and his research team have been studying virus clearance during MAb purification, as well as the isolation of reverse chimeric (i.e. murine) MAbs.

Jan 23: (A) Introduction to Course Objectives and (B) Topics/Overview of Monoclonal Antibody Process Development

Monoclonal antibodies (MAbs) are an important and growing class of therapeutic proteins. There are currently 21 MAbs on the market (and dozens more in clinical trials and at the research stage) with a wide variety of therapeutic indications. MAb process development and production methods will play a major role in the industry’s ability to effectively bring this large number of products to the market rapidly and economically. This lecture will provide an overview of the various key elements of MAb production including: genetically engineering cell lines to produce high levels of MAbs; designing nutritional media and optimizing bioreactor conditions that promote maximum cell growth and MAb production; scaling up bioreactor processes to pilot-scale and manufacturing-scale facilities; designing purification processes that maximize product yield while maintaining product quality; using bio-analytical methods to characterize key product attributes. After this lecture, students should understand the fundamentals of MAb process development and production, thereby preparing them for subsequent lectures in this field.

Reading Material:
- Chu and Robinson. "Industrial Choices for Protein Production by Large-Scale Cell Culture." Curr. Opin. Biotechnol. 2001, 12:180-187.
- Andersen and Krummen. "Recombinant Protein Expression for Therapeutic Applications." Curr. Opin. Biotechnol. 2002, 13:117-123.
- Shukla et al. "Downstream Processing of Monoclonal Antibodies--Application of Platform Approaches." J. Chromatogr. B. 2007, 848:28-39.

NO ASSIGNMENT

Instructors: Gregory Russotti/Bruno Marques

Jan 30: Mammalian Cell Culture Scale-Up for Monoclonal Antibody Production

Scale-up of monoclonal antibody cell culture processes to the pilot-scale, for the production of clinical material, and to the manufacturing scale, for commercial manufacture, will be discussed. When scaling up these bioreactor processes, challenges include: avoiding cell damage; providing adequate mixing; satisfying cellular oxygen demand; removing carbon dioxide from the bioreactor; and minimizing foam accumulation in the bioreactor—All while maintaining sufficient cell growth and productivity. Students will learn about the various approaches to scaling up agitation and aeration parameters, as well as the advantages and disadvantages of each with respect to scale-up challenges.

Reading Material:
- Chapter by Xie et al in Gene Transfer and Expression in Mammalian Cells.
- Mostafa and Gu. Biotech Prog. 2003, 19:45-51.
- Zhu et al. Biotech Prog. 2005, 21:70-77.

ASSIGNMENT:
CO2 transfer rate (CTR) out of a bioreactor is a function of mass transfer coefficient (kLa) and driving force (Cbulk minus C*), where Cbulk equals the concentration of CO2 in the bulk and C* equals the saturation concentration of CO2 based on the partial pressure of CO2 in the gas phase.  Discuss how these six parameters might affect CTR: (1) sparge rate, (2) agitator speed, (3) bubble size, (4) bicarbonate concentration, (5) impeller position, and (6) presence of antifoam.  Specifically, describe the physics behind how each of these might affect one or more of the following: kL, a, Cbulk, C*.

Instructor: Gregory Russotti

Feb 6: PEGylation of Proteins—Modification of Pharmacokinetic Properties

Although decisions about protein therapeutics are made primarily on the basis of biological activity, other properties such as pharmacokinetic behavior are critically important as well. This is especially true for relatively small protein therapeutics which may be cleared from the body too quickly to allow for effective therapy. One solution to this problem has been to attach a larger chemical moiety such as polyethylene glycol (PEG) to the protein to increase its half-life in circulation. The basis for this approach will be reviewed as well as how PEGylation fits in to a typical protein production process. In addition, other approaches to modifying the pharmacokinetics of proteins will be reviewed.

Reading Material:
- Fishburn CS. The pharmacology of PEGylation: Balancing PD with PK to generate novel therapeutics. J Pharm Sci. 2008 Jan 15.
- Harris JM, Chess RB. Effect of pegylation on pharmaceuticals. Nat Rev Drug Discov. 2003 Mar;2(3):214-21.

ASSIGNMENT:
You are the Head of the Health Authority in Monaco.  You have been approached by a company, Sino-Shiva, offering generic PEGylated interferon.  The cost savings would be tremendous and could offset recent losses in the casinos due to card counters.  Based on the two attached references, one related to Roche's Pegasys and the other on Schering-Plough's Peg-Intron, what process, analytical, and pre-clinical (e.g., animal PK, etc) questions would you ask related to the control of their product in order for it to be approved in Monaco.
Rerefence for Assignment:
- Foser S et al. Protein Expr Purif. 2003 Jul;30(1):78-87.
- Grace MJ et al. J Biol Chem. 2005 Feb 25;280(8):6327-36. Epub 2004 Dec 13.

Instructor: Eugene Schaefer, Sc.D., Director of Process Technology, Bristol-Myers Squibb

Gene Schaefer received his B.S. degree in Chemical Engineering from Princeton University, and his M.S. and Sc.D. degrees in Biochemical Engineering from M.I.T. His M.S. thesis, supervised by Professor C. L. Cooney, investigated the production of the enzyme maltase by yeast, and his doctoral thesis, supervised by Professor D.I.C. Wang, focused on the production of a bio-surfactant. Gene worked for Genzyme for three years, in Boston and the UK, developing processes for a number of protein and carbohydrate products. He then led groups in several areas including purification, fermentation development and molecular & cell biology development, within the Biotechnology Development Group at Schering Plough for over thirteen years. Gene has held the position of Director of Process Technology at Bristol-Myers Squibb for the last six years.

Feb 13: Cell Line Development for Monoclonal Antibody Production

Monoclonal antibodies (MAbs) represent a successful therapeutic area in terms of a growing number of products approved by the FDA and their clinical success. MAbs are produced by clonal cell lines and, as a result, the production of the cell lines themselves is a central process that will be the focus of this session.

The use of living cells as production factories has important implications unique to MAb production. Production of cell line begins with many millions of inherently genetically variable potential host cells into which cDNAs encoding MAbs are transfected. The diversity is further increased by variable transfection of the cells with significant consequences to their quality or performance. These include effects on growth rate and biomass potential, stability of expression, post-translation modification of MAb, and MAb yield, among other critical factors. The task in cell line production then becomes the isolation of those rare cells engineered such that all of these characteristics are optimized. Further, to ensure that a selected cell line performs as expected, the FDA enforces guidelines for cell line used in biologics production. These guidelines, as much as any technical issue in cell line production, need to be taken into account for a commercially viable outcome.

We will begin by discussing the most widely used and well established technologies for cell line production. Using these methods, it typically takes up to two years to obtain a cell line that may be scaled-up for the 10,000L production runs that are commonly used for sufficient yield. We will also cover some novel technologies developed to improve yield and/or timelines. This is a particularly timely discussion, as it happens that today’s marketplace predicts rapid growth in the MAb biologics sector, forcing a juxtaposition of the benefits and risks of existing and novel technologies. Another force that may have a great and immediate influence is the potential legalization of generic biologics. Interestingly, one of the issues that complicates this discussion is establishing exactly how a generics producer could match every quality of the cell line used by the original brand-name producers, given all the complexities in cell line development. We will discuss which aspects of cell line production are relevant in this context and, if resolved, how the advent of generic biologics may have its own mark on which technologies may be favored.

Reading Material
- Animal cell cultures: recent achievements and perspectives in the production of biopharmaceuticals. Appl Microbiol Biotechnol 68, 283 – 291 (2005)
- Production of recombinant protein therapeutics in cultivated mammalian cells. Nature Biotechnology  22, 1393 - 1398 (2004)
- Evolution of cell-based reagents provision. Drug Discovery Today 12 820 – 825 (2007)

ASSIGNMENT:
1) What are the critical qualities that a Mab production cell line must meet for optimal efficiency of Mab production, and how can a sub-optimal cell line affect the overall process?
2) The paper on the evolution of cell-based reagent provision considers the use and limits of cell lines in drug discovery (as opposed to Mab production). Which points raised in this discussion are also relevant to cell line production for biologics?

Instructor: Kambiz Shekdar, Ph.D., Chief Scientific Officer, Chromocell Corporation

Kambiz Shekdar is a co-founder of Chromocell Corporation and has served as its Chief Scientific Officer since its inception in 2003. From 1992 to 1995, Kambiz attended Rutgers College where he received his B.A. in Biology. He was a Henry Rutgers Scholar in the Department of Biology at Rutgers College in 1995. From 1995 to 2003, Kambiz was a Graduate Fellow in the laboratory of Günter Blobel at The Rockefeller University in New York City where he obtained his Ph.D. His doctoral thesis work resulted in the biochemical purification and identification of a network of intra-nuclear fibers from rat liver nuclei which may represent a novel intra-nuclear transport or organizational scaffolding. During the course of this work, Kambiz had to create many stable cell lines. This normally required extensive manual tissue culture work and, as a result, Kambiz and Gunter co-invented a technology that automates cell engineering. At Chromocell, Kambiz has worked closely with CEO Christian Kopfli to establish the company. During the start-up phase, this included bench work in addition to coordination of the initial research program and collaboration projects. The company rapidly established itself and continues to grow at a fast pace; Kambiz currently oversees the scientific management of the various projects and scientifically contributes to corporate development of the company.

Feb 20: DNA Vaccines Product Development—From the Bench to the Clinic

Vaccines have played a pivotal role in increasing life expectancy worldwide and, in particular, the developed world has seen life expectancy increase by over 30 years in the last century. Indeed, mortality from the nine major vaccine preventable diseases of the 19th century has decreased by over 99.99%. While conventional vaccines (live, live-attenuated or inactivated) have held the predominant position amongst licensed products, they have often been plagued with manufacturing, potency, and safety issues making the search and development of newer vaccine platforms an attractive area for innovation. The key drivers being: ease of production, reduction in cost of goods, improving lot-to-lot consistency, stimulation of humoral and cellular immunity, improved efficacy, ease of delivery, better safety/toxicity profile, dose-sparing regimens, and rapid response strategies.

Plasmid DNA based vaccines and vaccines using viral vectors (adenovirus, MVA, and others) hold many of the attributes desirable in a vaccine candidate. While DNA-based vaccines have great potential, the best current DNA vaccines induce only weak cellular immunity in humans and are mainly being tested in the clinic as priming modalities that only weakly improve subsequent recombinant viral boosting. This lecture will focus on introducing students to the attributes that make DNA vaccines attractive for vaccine product development. We will start by discussing factors to consider while designing vectors, antigens and host strains. We will discuss current manufacturing processes for DNA and the peculiarities of DNA that makes process development and scale-up a challenge. We will touch upon the regulatory environment for DNA-based biologics. Lastly, we will discuss strategies to improve the immune potency of DNA vaccines and discuss cases that have generated widespread excitement (and controversy) in the community for the role of T-cell mediated vaccines.

Reading Material
- Liu M. et al. 2006. Human Gene Therapy. 17:1051-1061.
- Cristillo A.D. et al. 2008. Biochem. and Biophys. Res. Commun. 366:29-35.
- Lowrie D. 2006. Vaccine. 24:1983-1989.

ASSIGNMENT (note: HW question number 2 is a bonus/optional question):
1) What are some of the advantages afforded by DNA as a vaccination methodology?
2) BONUS QUESTION/OPTIONAL: While there are numerous sources of obtaining “research grade” plasmids, there are only a few manufacturers focusing on cGMP plasmid manufacturing worldwide. Using the internet, identify some of the lead cGMP manufacturers of plasmid DNA. What is the total installed capacity based on fermenter size (L) at these manufacturers? Provide an estimate of the worldwide cGMP capacity for plasmid DNA?

Instructor: Niranjan Y. Sardesai, Ph.D., Sr. Vice-President, Research & Development, VGX Pharmaceuticals, Inc.

Niranjan Sardesai is currently the Senior Vice President of Research and Development at VGX Pharmaceuticals, headquartered in Blue Bell, PA. VGX is a clinical-stage biopharmaceutical company engaged in the development of novel drugs, and DNA-based vaccines and therapeutics, for major infectious diseases and cancer. Dr. Sardesai is responsible for formulating and directing the company’s development strategy, as well as for directing the pre-clinical research programs. He also has direct oversight and P&L responsibility over the company’s state-of-the-art cGMP manufacturing facility for plasmid DNA located in The Woodlands, TX. Previously, Dr. Sardesai served as VGX’s Vice President of Product Development. He is an experienced veteran of the pharmaceutical industry, with a special focus on R&D and Management of Technology. Prior to VGX, Dr. Sardesai served as Founder and President of NVision Consulting Inc., a strategy consulting firm focused on entrepreneurial companies in the biopharma/life-science industry, and as Director of R&D at Fujirebio Diagnostics, Inc. At FDI, Dr. Sardesai oversaw all aspects of R&D activities and expansion of the oncology portfolio. He was responsible for establishing the R&D division following their acquisition from Centocor and for rebuilding the product pipeline. Products developed under his leadership include new-to-the-world diagnostic tests for mesothelioma (MESOMARK™), bladder cancer, and a multi-marker test for ovarian cancer. Dr. Sardesai also served as Senior Scientist at IGEN International, Inc., where his group helped develop a high-throughput screening platform for drug-discovery.

Dr. Sardesai received a Ph.D. in Chemistry from the California Institute of Technology, and an MBA in Entrepreneurship and Finance from the Wharton School of the University of Pennsylvania. He completed Fellowships at the Scripps Research Institute and at the Massachusetts Institute of Technology. Dr. Sardesai received his M. Sc. in Chemistry (5-year Integrated Program) from the Indian Institute of Technology.

Feb 27: Large-Scale Bioreactor Design and Application

Bioreactors are the most complex upstream bioprocess equipment item in the biotechnology and pharmaceutical manufacturing industries. They are the “factories” in which cultivation of cells to high densities permit the expression of useful proteins derived from recombinant DNA techniques. Drug companies that are in late stage phase III clinical trials or about to launch an FDA approved drug require facilities that can produce sufficient quantities of their product to meet market demands. A train of bioreactors having increasing vessel size is necessary for large scale production of modern therapeutic proteins, viruses, vaccines, monoclonal antibodies and a host of other biologically derived products in kilogram quantities. Scaling-up microbial fermentors and cell culture bioreactors can present challenges to the process and equipment designer. A thorough understanding of the inter-relationships between vessel geometry, aeration, agitation (mixing), oxygen transfer, and heat transfer are necessary to fully appreciate the scale-up methodology from laboratory to full-scale production. This lecture will explore OUR (Oxygen Uptake Requirements), agitator power needs, heat transfer demands, and aeration using air and / or oxygen. Control of critical parameters will be discussed as they relate to the real time measurement of gas flows, pressure, temperature, agitator speed, pH, DO (Dissolved Oxygen), and off-line measurements.

Reading Material:
- Stadler E, 1998. Dual Purpose Fermentator and Bioreactor? A Capital Quandry! Pharm. Eng., Vol 18(3)

ASSIGNMENT:
Inlet and exhaust gas management to support metabolism for a cell culture bioreactor varies considerably from a microbial fermentor. List and compare the key elements that are different and identify the affect of size on the hardware chosen to control variables such as sparge gas distribution, mass flow off gas, pressure, and dissolved oxygen. All of these contribute to success for supporting optimum Oxygen Transfer Rate (OTR) for a good Kla value.

Instructor: Ernest L. Stadler, P.E., Director Custom BioProcess Systems, Sartorius Stedim Biotech

Ernest L. Stadler is Director of Custom Bioprocess Systems at Sartorius Stedim Biotech, a manufacturer of fermentors, bioreactors, automation software / hardware, and other related bioprocess laboratory and custom large scale equipment. A member of ASME, ISPE, and PDA, Mr. Stadler is a registered Professional Engineer in PA and NJ, holding a BS in Mechanical Engineering from NJIT and has done graduate studies at Lehigh University in biotechnology. Mr. Stadler has broad based expertise in automation, process and mechanical design for a wide range of Bioprocess equipment having served the biotechnology industry for over 19 years. He is a frequent speaker, teacher, and workshop leader on the design and application of fermentors and bioreactors, particularly relating to biotechnology process scale-up and optimizing performance. Ernest served as Course Director, Lecturer and Workshop Leader for over 12 years with the ASME BioProcessing Technology Seminar series. He has authored articles and papers and was awarded the 1998 Article of the Year by Pharmaceutical Engineering Magazine, a publication of the ISPE. He has served the past 3 years on the Board of Directors for the SBP being a founding member and is the current SBP Treasurer. Mr. Stadler can be reached via email at ernest.stadler@sartorius.com

Mar 5: Recovery Process Development and Validation for a Therapeutic Monoclonal Antibody

Monoclonal antibodies (MAb) represent the largest class of protein therapeutics in the biotechnology industry. While all MAbs share many characteristics, each MAb has a unique combination of biochemical properties that can be exploited or that must be overcome for purification. Successful development of a MAb purification process also requires consideration of product quality, economics, manufacturing constraints, regulatory and product safety issues, and speed of development. These factors must be continually evaluated as the process is transferred to a pilot manufacturing facility and, ultimately, to a commercial manufacturing facility. Prior to obtaining a license to launch the MAb commercially, a process validation package must be prepared to demonstrate that the manufacturing process will consistently perform as the manufacturer claims. Process development, and validation strategies and alternatives will be discussed using illustrative examples.

REQUIRED:
- ICH Guideline Q5E, “International Conference on Harmonisation (ICH); Guidance for Industry: Q5E Comparability of Biotechnological/Biological Products Subject to Changes in Their Manufacturing Process” available at http://www.fda.gov/CbER/ich/ichguid.htm 
- FDA Guidance for Industry, “Comparability Protocols - Chemistry, Manufacturing, and Controls Information” available at http://www.fda.gov/cber/gdlns/cmprprot.htm

BACKGROUND:
- Purification Tools for Monoclonal Antibodies, by P. Gagnon, Chapters 1, 3-7, 9
- Protein Purification, Principles and Practice, by R. Scopes, Section 1.3, Chapters 5-6
- Guide to Protein Purification, by M.P. Deutscher. Section VII

ASSIGNMENT:
Summarize when manufacturers are expected to demonstrate mAb drug substance comparability.  Describe how a manufacturer might determine what information is required to demonstrate comparability.

Instructor: William K. Wang, Ph.D., Director, Process Biochemistry, MedImmune

William K. Wang is currently Director, Process Biochemistry at MedImmune, a biopharmaceutical company in Gaithersburg, MD. Bill leads a team of scientists and engineers responsible for process development, technology transfer, and clinical and commercial manufacturing support. Prior to joining MedImmune in 2007, Bill supported purification and filling operations, and process transfer for therapeutic proteins at Biogen Idec’s Cambridge, MA commercial and clinical manufacturing facility. Prior to joining Biogen Idec in 2002, Bill developed and transferred purification processes for clinical manufacturing of therapeutic proteins at GlaxoSmithKline. He completed his M.S. and Ph.D. in Chemical Engineering from the University of Rochester. His doctoral research focused on the cloning and expression of the gene coding for a Clostridium thermocellum cellulase. Bill completed his B.S.E. in Chemical Engineering at the University of Pennsylvania. He was the editor of the book, Membrane Separations in Biotechnology: Second Edition, Revised and Expanded and is an author on over 25 articles, chapters, or abstracts for presentations. Recently, he was a Program Chair for the Division of Biochemical Technology at the 2007 ACS National Meeting.

Mar 12: Live Virus Vaccine Process Development

Viral vaccines are an important class of therapeutics in the pharmaceutical industry. A production challenge in live virus vaccine manufacturing is the large-scale cultivation of viruses. Viruses, by themselves, are non-replicating organisms and, therefore, need a host in order to replicate. Typical hosts include in vivo, in ovo (egg cultivation), and in vitro systems. Primary cells were the traditional cell type for in vitro systems but, more recently, human diploid cells and continuous mammalian cell lines have been developed for viral cultivation. The cells used in these in vitro systems are anchorage-dependent; that is, they require an attachment surface in order to grow. Various cell culture configurations for anchorage-dependent cells will be compared, and the advantages and disadvantages of these configurations—with respect to scale-up, economics, process robustness and control—will be discussed.

Reading Material::
-Junker BH, Wu F, Wang S, Waterbury J, Hunt G, Hennessey J, Aunins J, Lewis J, Silberklang M, Buckland BC. Evaluation of a microcarrier process for large-scale cultivation of attenuated hepatitis A. Cytotechnology. 1992;9(1-3):173-87.
- Aunins et al., Vaccine Production
- Cell factories

ASSIGNMENT:
Compare the use of microcarrier culture in a bioreactor to static culture using a disposable reactor technology such as Nunc Cell Factories for the production of a live virus vaccine. Discuss the advantages and disadvantages of each.

Instructor: Gregory Russotti

Mar 26: Technology Transfer of a Biological Product from Process Development to Manufacturing
PLEASE NOTE: Speaker, reading material and assignment has changed.

As a biological or pharmaceutical product moves through a company’s pipeline, it will undergo process development, scale-up, and testing through Phase I and Phase II clinical trials. If the product proves successful from a clinical stand-point, the financial feasibility of manufacturing the product on full-scale will be evaluated by the company. Factors such as manufacturing location, manufacturing costs, expected time to licensure, and current anticipated sales, will factor into the Net Present Value (NPV) assigned to the product. Based on proven clinical results and a positive NPV, the process technology will be transferred into manufacturing in preparation for Phase III clinical trials and product licensure. The steps associated with a successful technology transfer will be discussed, including facility design fundamentals and start-up, process demonstration, and product licensure.

Reading Material:
- Goochee, C. The roles of a process development group in biopharmaceutical process startup. 2002. 38:63-76.

ASSIGNMENT:
The assigned reading focuses on the activities that a process development group- and to a lesser extent manufacturing groups- should perform during a process transfer and in the months leading up to it. Please discuss what the process development and manufacturing groups might do 1-5 years prior to the launch of a commercial manufacturing facility to improve the probability of a successful plant start-up..

Bonus question: In a single word or with a short phrase, please identify what you think is the most important thing that the process development and manufacturing groups can do before and during a process transfer to improve the probability of success.

Instructor: Gregory Russotti

Apr 2: Technology Evaluation and Process Development Strategy for the Production of Therapeutic Proteins

This session will address the available technologies for the large-scale production of biopharmaceuticals. The relatively high approval rate for high-demand and high-dose biological products has raised concerns regarding production capacity, reliability on the production technology, and the economics for this type of product. This class will review current literature on process scale-up and technology assessment. The class will focus on the strategy used to reconcile process development timelines and appropriate production scale, with the uncertainties associated with product development and investments required to support it.

Instructor: Marco A. Cacciuttolo, Ph.D., President and CEO, PERCIVIA LLC.

Dr. Cacciuttolo is the Chief Executive Officer of PERCIVIA LLC, the PER.C6 Development Center located in Cambridge, Massachusetts. This center is a joint venture between Crucell, B.V. and DSM Biologics, B.V. Previously, Dr. Cacciuttolo was Vice President of Technical Operations at Medarex, Inc., a biotechnology company with headquarters in Princeton, New Jersey. In that role, Dr. Cacciuttolo was responsible for process development, analytical sciences, quality control, GMP manufacturing, validation, materials management, and the management of outsourcing activities. Dr. Cacciuttolo was previously Manager, Cell Culture Manufacturing at MedImmune, Inc., Frederick, Maryland. In this role, Dr. Cacciuttolo was responsible for the manufacturing of Synagis, the first approved monoclonal antibody against an infectious agent. He also participated in the commissioning and licensure of the facility to manufacture Synagis. Prior to this assignment, Dr. Cacciuttolo was a process development engineer in Cell Culture Development at MedImmune, Inc., Gaithersburg, Maryland. Dr. Cacciuttolo received his Ph.D. in Biochemical Engineering from the University of Maryland, Baltimore County and his B.S. in Biochemical Engineering from the Catholic University at Valparaiso, Chile.

Reading Material:
- Cacciuttolo, M.A., and Arunakumari, A. 2006, "Scale-up Considerations for Biotechnology-Derived Products", in Pharmaceutical Process Scale-Up, ed. Michael Levin, 2nd edition, Taylor & Francis, ch. 5.
- Farid, SS., 2007, Process Economics of Industrial Monoclonal Antibody Manufacture, J. Chrom. B., 848:8-18.
- Kelley, B., 2007, Very Large Scale Antibody Purification: The Case for Conventional Unit Operations, Biotechnol. Prog., 23:995-1008.

ASSIGNMENT:
Background: The biotechnology products can be classified into the following major categories:
1. Brand new, first line therapy (no previous medicine/therapy available)
2. Brand new, first in class (a new approach to treat a disease. That is, replacing standard therapy for a better approach, less side effects, lower dose, better survival, easier to use, etc.)
3. "Bio-better": same validated target than brand new, better economics (better molecule, process, clinical protocol, delivery, or combinations thereof)
4. Biosimilar ("same" everything: molecule, clinical protocol, delivery, etc)

Question: you are a new player in the industry and area about to decide which product to develop. Select one product from the above list. Explain the rational for choosing both category of product to develop and the manufacturing platform to produce it.

Apr 9: Challenges and Strategies in Cell Therapy and Tissue Engineering Bioprocess Development

When living cells are used as therapeutics, the challenges of meeting FDA safety, efficacy and manufacturing standards are amplified by the inherent variability of the product. The additional challenges posed by process and distribution economics have kept the number of such therapeutics that reach wide clinical distribution quite small. Notwithstanding these obstacles, the great promise of this field as a component of the new clinical paradigm of regenerative medicine has stimulated continuing efforts to bring new such medical devices into clinical trials.

This session will focus on cell based therapies that rely on normal cells recovered from human tissues. The emphasis will be on practical approaches to address FDA issues, from tissue procurement through cell line and final product characterization; and process economics issues, from pilot scale through commercial launch. A case study will be included, of the product OrCel®, produced by Forticell Bioscience (formerly Ortec International). OrCel is a tissue engineered skin substitute in the form of a cryo-preserved bilayered cellular matrix in which normal human donor epidermal keratinocytes and dermal fibroblasts are cultured. Forticell entered into an agreement with a contract manufacturer for the late stage clinical trial supply and commercial manufacture of OrCel. The ensuing technology transfer and production scale up will be discussed.

Reading Material:
- Eisenbud E, Huang NF, Luke S, Silberklang M. Skin Substitutes and Wound Healing: Current Status and Challenges. Wounds 2004;16(1):2–17
- Applegate DR, Liu K, Mansbridge J. Practical Considerations for Large-Scale Cryopreservation of a Tissue Engineered Human Dermal Replacement
- Parenteau NL, Bilbo P, Nolte CJ, Mason VS, Rosenberg M. The organotypic culture of human skin keratinocytes and fibroblasts to achieve form and function. Cytotechnology. 1992;9(1-3):163-71.

Supplemental Reading:
- Ehrenreich M, Ruszczak Z. Update on tissue-engineered biological dressings. Tissue Eng. 2006 Sep;12(9):2407-24.
- Auger FA, Berthod F, Moulin V, Pouliot R, Germain L. Tissue-engineered skin substitutes: from in vitro constructs to in vivo applications. Biotechnol Appl Biochem. 2004 Jun;39(Pt 3):263-75. Review.

ASSIGNMENT:
1. You have come up with a brilliant engineering coup and have found a better and more cost effective way to produce a previously FDA licensed “Skin Substitute” product. Your objective is to convince FDA to approve the new process without demanding an entirely new clinical trial. Using the two known licensed products, Apligraf® and Dermagraft®, describe, for each one, how you might go about demonstrating the bio-equivalence of product made through your new process to the original licensed product. Keep in mind both safety and performance issues.

2. If current product sales are 50,000 units per year at $1,000/unit, and sales are growing at 25% per year – assuming a constant annual sales rate of growth, what cost saving per unit (expressed to the nearest dollar) would be necessary to recoup a $10 million overall project investment within 18 months of launching the new process? Within 24 months? Within 36 months?

Instructor: Melvin Silberklang, Ph.D., Chief Scientific Officer and Vice President, Research and Development, Forticell Bioscience

Mel Silberklang is currently Chief Scientific Officer and Vice President of Research and Development at Forticell Bioscience (formerly known as Ortec International). He has over 26 years of experience in the Pharmaceutical (Merck Research Laboratories, 1981-1993) and Biotechnology (Enzon, Inc., 1993-1995; Ortec International, Inc., 1995-2007) industries and two years of prior academic experience on the research faculty of the University of California at San Francisco. His background in biotechnology is very broad, spanning the gamut from discovery research through product development and production. He has worked on: gene cloning and synthesis; protein and antibody engineering; high-level animal cell expression; live and subunit viral vaccine development and production; primary cell and stem cell isolation, characterization and culture; process scale-up and development and GMP production of proteins, monoclonal antibodies, vaccines, cell therapies, stem cell therapies, tissue-engineered three-dimensional tissue-like cell cultures, biomaterial matrices and scaffolds. Since assuming the position of Vice President for Research and Development at Ortec (1996), he has been involved in every step of a tissue engineering project to develop a cultured, bi-layered cellular matrix as a wound healing product, trademarked OrCel. The program spans: high-grade (bovine) collagen procurement, testing, and processing; human tissue (foreskin) procurement and processing; primary skin cell line establishment, expansion, cryo-preservation, safety and efficacy testing; final product process development, technology transfer, production, Quality Control, packaging, cryo-preservation and shipment. The OrCel® product has gotten two FDA approvals and is presently undergoing FDA PMA review of an advanced cryo-preserved form. Dr. Silberklang is presently also directing research into optimization of autologous adult stem cell isolation, characterization, expansion, differentiation, and therapeutic delivery using natural and synthetic biomaterial matrices.

Apr 16: Standard Oil and the Vertically Integrated Pharma Model: Repeating History

In 1911, the U.S. government broke up Standard Oil, citing its monopoly position in all aspects of energy production ("vertically integrated," from exploring/drilling to retail delivery of product). In a similar action, AT&T (responsible for everything from handset production to undersea cable maintenance) was broken up in 1984. In both cases, a credible argument can be made for a wave of innovation and value-creation following on the heels of the break-ups. The pharmaceutical industry has been traditionally vertically integrated, with only the clinical trial aspects obligatorily contracted out. Big Pharma, perhaps inspired in the 1980s by the hope that genomics would provider a "unified field" approach to human health, began a further consolidation, leading to the mega-companies of today. With genomics, and other expensive front-end technologies, including high-throughput screening, it seemed to make sense to grow, amortizing these costs over a larger base.

While it is unclear that these changes causally lead to the current "innovation crisis," Pharma has recently started to shift business models, moving away from vertical integration towards a model of "open innovation." This has implications for all aspects of the business, and the science and technology it supports. We will discuss the underlying observations, and the implications for employment in the industry. Also to be considered is outsourcing, a.k.a. the "Asia Model."

Reading Material:
Note, please read ALL the paper/article ABSTRACTS, then ONLY 2-3 of the ARTICLES. "Full marks" for partial answers which reflect the questions posed in the subset of papers chosen, when applied to the questions below. As an FYI, I am willing to shamelessly take your ideas to help save my industry.
- A bitter pill for Big Pharma - Los Angeles Times
- The Impact of Genomics on Drug Discovery
- Stochasticity and Cell Fate
- Type 2 Diabetes May Be Caused by Intestinal Dysfunction
- Modeling network dynamics: the lac operon, a case study
- Sequence- and target-independent angiogenesis suppression by siRNA via TLR3
- Metabonomic Identification of Two Distinct Phenotypes in Sprague-Dawley (Crl:CD(SD)) Rats
- All in the Stroma: Cancer’s Cosa Nostra
- Tiny Gene Variations Can Even Alter Effects of the Pills We Take
- HIV Gets By With a Lot of Help From Human Host
- Reversible Compartmentalization of de Novo Purine Biosynthetic Complexes in Living Cells

ASSIGNMENT:
The lecture will start with the assignment of blame for the current state of Pharma Research and Development, the underlying theme will start to address how to approach large and complex problems with minimal data.

1) The development of a new pharmaceutical can take 15 years and ~$1B, what kind of assumptions are necessary to justify the investment?

2) Engineering generally assumes a reasonable understanding of the "properties of materials", can it be usefully applied to complex systems (life)?

3) From the readings, it becomes evident that all medicine is "personalized" (because each patient is absolutely individual). How would you approach true personalized medicine (right chemical into the right body)? Is it conceivable that patients can be placed into few enough "classes" to allow the production of drugs optimized for efficacy and safety (at least for the bulk of the population)? Are there any other approaches to the problem that might not require a 15-year lead time (and associated guesswork)?

Instructor: Robert A. Zivin, Ph.D., Corporate Director, Johnson & Johnson Corporate Office of Science and Technology

Bob Zivin received his B.S. in Biology at Northern Illinois University and his PhD. in Microbiology at the University of Chicago. His thesis work was directed toward the molecular genetic characterization of an atypical bacteriophage. After a post-doc at the National Cancer Institute, Bob spent several years at the Merck Research Laboratories in West Point, PA, working on the cloning and bacterial production of therapeutic peptides. Bob has spent over eighteen years with Johnson & Johnson where, among other responsibilities, he led the Antibody Humanization effort at the R.W. Johnson Pharmaceutical Research Institute. In 1995, Bob established and headed the Department of Exploratory Technology at RWJ PRI (now J&J Pharmaceutical Research and Development, L.L.C.), working on the application of new technologies to the drug discovery process. Since November 2003, he has been a member of the J&J Corporate Office of Science and Technology.

Apr 23: Cellular Therapeutics: Accomplishments, Promises, and Challenges

There has been limited but successful use of cells in medicine, and there is great promise for additional cell-based therapeutic applications. Numerous challenges related to medicine and biology, safety/regulatory, manufacturing, and business must be overcome during further development of cell therapies. As the ability to address these issues hinges on all aspects of design, the bioengineering of a product is critical to its success. This session will focus on the current state of the cell therapy field, and engineering approaches to dealing with challenges. Among the challenges that will be discussed are cell sourcing, matching, scale-up, cell banking, testing, and storage. This lecture will also provide an overview of developments at some notable companies pursuing cell therapies.

Reading Material:
- FDA Draft Guidance for Reviewers: Instructions and Template for Chemistry, Manufacturing, and Control (CMC) Reviewers of Human Somatic Cell Therapy Investigational New Drug Applications (INDs)
- Will regulation be the death of cell therapy in the United States? By DA Gastineau

ASSIGNMENT:
Would you be willing to be treated with a cell therapy that did not conform to all of the requirements described in the FDA guidance?  Which, if any, of the requirements would you be willing to overlook?  How does your decision depend on the disease for which you might be treated?

Instructor: Thomas A. Brieva, Ph.D., Senior Research Scientist, Cellular Process Development, Celgene Cellular Therapeutics

Tom Brieva received a B.S. in Chemical Engineering and a B.A. in Biological Sciences from Rutgers University. His Ph.D., also from Rutgers, combined Chemical Engineering and Cell and Developmental Biology in an Interdisciplinary Ph.D. program aimed at training useful for Tissue Engineering. His Ph.D. research involved engineering hepatocyte growth and function via cell-cell adhesion receptors under the direction of Dr. Prabhas Moghe. Tom also completed two NIH-sponsored training programs: the Biotechnology Training Program and the Molecular Biophysics Training Program. Following post-doctoral research on cell targeted nanocarrier-based drug delivery in Dr. Joachim Kohn’s lab at the NJ Center for Biomaterials at Rutgers, Tom began his current tenure at Celgene. He presently pursues processes for the isolation, expansion, and formulation of therapeutic placental stem cells in a safe, robust, reproducible, controlled, and cost-effective manner. Collectively, Tom’s work throughout his career has focused on engineering cells via culture processes, cues, or small molecules.

Apr 30: Development and Commercialization of a Regenerative Tissue Matrix for Soft Tissue Repair and Replacement

Regenerative Medicine is a relatively new field that encompasses the disciplines of cell biology, extra cellular matrix biochemistry, bioengineering, transplantation, immunology, and tissue engineering, all focused on harnessing and directing the body to regenerate new tissues and organs. A number of approaches are under investigation in many academic and industry programs and include: Replacement of tissue with entirely synthetic materials constructed ex vivo; Functional restoration with constructs that comprise both synthetic and cellular components; Scaffolds that revitalize in vivo and transition to normal tissue; Combinations of temporary scaffolds (natural or synthetic) with cellular components; and Cellular therapies, including stem cells and genetically modified cells. At LifeCell Corporation we have developed and commercialized extra cellular matrix-based products for soft tissue repair and reconstruction. The key to success of the technology has been driven by recognition of the importance of preserving the native structure and composition of the extra cellular matrix. When implanted in the body, the intact scaffold is rapidly vascularized and populated by host cells with minimal signs of inflammation. Positive recognition by the body is key to remodeling to native tissue and long term efficacy; and clearly distinguishes this material from other biologic or synthetic materials that elicit immune rejection and subsequent resorption or encapsulation. Development of an intact, sterile, regenerative tissue matrix derived from an animal source will be described. This will include procurement from a controlled animal herd, processing methods that will scale to meet demand, overcoming graft rejection in animal to human xeno-transplantation, long-term room temperature preservation of a complex biologic, sterilization without degrading or cross linking extra cellular matrix components, and regulatory considerations in choosing the product development path. The preservation of key matrix components was verified by in vitro and in vivo testing, and efficacy assessed by evaluation in a number of animal models including non-human primates. The pathway to final clearance by the FDA as a 510(k) medical device will be described, as well as early clinical evaluation in hernia repair and post-mastectomy breast reconstruction. This lecture will touch on the multidisciplinary approach required to successfully address unmet needs in regenerative medicine, encompassing tissue structure and function, immunology of transplantation, identification and development of appropriate animal models, and issues related to regulatory requirements in developing a medical device.

Reading Material
Required:
- Guidance for the Preparation of a Premarket Notification Application for a Surgical Mesh
- Cellular and Molecular Dynamics in the Foreign Body Reaction
- Extracellular Wound Matrices: A Novel Regenerative Tissue Matrix (RTM) Technology for Connective Tissue Reconstruction
- Xenotransplantation: current status and a perspective on the future

Additional/Optional:
- Collagen Family of Proteins
- Source Animal, Product, Preclinical, and Clinical Issues Concerning the Use of Xenotransplantation Products in Humans

Instructor: David J. McQuillan, Ph.D., Vice-President Research, LifeCell Corporation

David McQuillan is Vice-President for Research at LifeCell Corporation and has been in that position since 2002. From 2000 to 2002 he was Director of Research at LifeCell. Prior to LifeCell, David spent 20 years in academia studying extra cellular matrix biology, the role in biosynthesis of orthopedic tissues, structure and function of extra cellular proteoglycans and collagens, and the role in wound healing and tissue repair. David received his BSc and PhD degrees from Monash University, Australia in Connective Tissue Research. He did postdoctoral work at the National Institute of Dental Research at NIH from 1985-1989, studying carbohydrate chemistry. He was a NH&MRC Research Fellow in the Orthopedic Research Unit, University of Melbourne 1990-1994, studying heritable diseases of connective tissue metabolism and studying interactions of matrix molecules by over-expression of recombinant glycoproteins. In 1994 David was recruited to the Institute of Biosciences and Technology, Texas A&M University where he managed an NIH-funded program studying the function of extra cellular matrix molecules.