Cell Culture Bioprocess Learnings: Past successes and future challenges

Therapeutic proteins, vaccines and – further away – cell and gene therapies are considered essential to find efficacious interventions and preventive measures to a rather long list of prioritized needs in healthcare. The challenge is developing into a ‘tsunami’ of sorts as the global demographic transition causes populations to age, dramatically accelerating the growth of non-communicable disease (NCD) in all parts of the world. India and China are the markets with the largest patient population for almost any NCD, or they are going to be in that position soon. Competition in this field is ramping up with an intensity that was previously unknown. Not only are there typically five to ten molecules from western originator companies for the same medical indication, but now there is finally also a huge wave of biosimilars entering the global markets. Asia is a hotspot in three ways: as the largest future market, as a location for manufacturing, and as an area with growing significance for the development of biologics. These developments all focus management teams onto financial performance of their businesses, namely via pricing of drugs and vaccines and cost of operations, not the least manufacturing cost. Different approaches to think about setting up manufacturing technologies such as continuous processing and incorporation of single-use equipment into routine large scale production are all part of this economic improvement discussion. Individualized therapies at the extreme for just one patient are an exciting medical route but process scientists and regulatory affairs must have nightmares over cost and approval challenges. Healthcare payers are already alarmed looking at annual treatment costs between $10’s to 100’s of thousands per patient with current therapies produced with the best of todays’ manufacturing tools. Regulators find proteins complex enough, cells must look like ‘mission impossible’ to them with today’s approaches. The discussion about future strategies and technical approaches goes far beyond the science and the engineering of bioprocesses. This presentation aims to describe the future scenario with its challenges, extract what was truly successful in the past, but then also de-mystify some of the hype topics in technology often considered as the holy grail of future manufacturing. Finally, there will be more questions than answers and these shall be the framework for this conference the ones to come

Individualized Biventricular Epicardial Augmentation Technology in a Drug-Induced Porcine Failing Heart Model

For treatment of advanced heart failure, current strategies include cardiac transplantation or blood-contacting pump technology associated with complications, including stroke and bleeding. This study investigated an individualized biventricular epicardial augmentation technology in a drug-induced porcine failing heart model. A total of 11 pigs were used, for the assessment of hemodynamics and cardiac function under various conditions of support pressures and support durations (n = 4), to assess device positioning and function by in vivo computer tomographic imaging (n = 3) and to investigate a minimally invasive implantation on the beating heart (n = 4). Support pressures of 20-80 mmHg gradually augmented cardiac function parameters in this animal model as indicated by increased left ventricular stroke volume, end-systolic pressures, and decreased end-diastolic pressures. Strong evidence was found regarding the necessity of mechanical synchronization of support end with the isovolumetric relaxation phase of the heart. In addition, the customized, self-expandable implant enabled a marker-guided minimally invasive implantation through a 4cm skin incision using fluoroscopy. Correct positioning was confirmed in computer tomographic images. Continued long-term survival investigations will deliver preclinical evidence for further development of this concept

Engineering Translation in Mammalian Cell Factories to Increase Protein Yield: The Unexpected Use of Long Non-Coding SINEUP RNAs

Mammalian cells are an indispensable tool for the production of recombinant proteins in contexts where function depends on post-translational modifications. Among them, Chinese Hamster Ovary (CHO) cells are the primary factories for the production of therapeutic proteins, including monoclonal antibodies (MAbs). To improve expression and stability, several methodologies have been adopted, including methods based on media formulation, selective pressure and cell- or vector engineering. This review presents current approaches aimed at improving mammalian cell factories that are based on the enhancement of translation. Among well-established techniques (codon optimization and improvement of mRNA secondary structure), we describe SINEUPs, a family of antisense long non-coding RNAs that are able to increase translation of partially overlapping protein-coding mRNAs. By exploiting their modular structure, SINEUP molecules can be designed to target virtually any mRNA of interest, and thus to increase the production of secreted proteins. Thus, synthetic SINEUPs represent a new versatile tool to improve the production of secreted proteins in biomanufacturing processes. \ua9 2016 The Author