Posters

High Throughput Development of Non-Protein A Monoclonal Antibody Purification Process using Mini-Columns and Bio-Layer Interferometry

High throughput (HT) sorbent screening is widely used for developing purification process saving time and sample volume. This study describes the development of a non-protein A purification process for a monoclonal antibody (MAb) expressed in a CHO cell supernatant. For the capture step, 54 process conditions were screened in three days using 200 µL mini-columns packed with five different sorbents. Capacity and yield were evaluated using Bio-Layer Interferometry (BLI) with protein A biosensors and MAb purity, using SEC-HPLC. A 340-fold scale-up of…

Efficient Strategies to Deliver Reliable and High Quality Biomanufacturing Processes Through Optimization of Cell Culture Media, Feeds, and Process Parameters

Development of a reliable and high quality biomanufacturing process for recombinant CHO cell lines presents challenges from diverse nutritional requirements observed with different clonally derived cell lines, and variable response to process operating ranges. To address these challenges, we will discuss efficient approaches to optimize CHO cell culture media, feeds, and process parameters within FUJIFILM Diosynth Biotechnologies (FDB). Also discussed is FDB’s new ApolloTM CHO expression system, a cornerstone of process scale-up efficiency offering robust gene expression in a platform…

Evaluation of Rapid Quantitation Methods for Titer, HCP and ProA By Gyrolab Platform During In-Process Development

Getting biopharmaceuticals through development and into clinical proof-of-concept fast and efficiently is critical for success in our industry. Therefore, high-throughput upstream and downstream process development approaches are increasingly being implemented. Hence, innovative and high-throughput analytical technologies are needed to support rapid process development. A novel automated analytical platform-Gyrolab xP workstation was investigated for the analysis of IgG titer, Host Cell Protein (HCP) and leached ProA during in-process development. The assays are automated within a Gyrolab compact disc (CD) containing affinity-capture…

Paired ADCC Reporter Bioassays Enable Differentiation of Antibody Fc Effector Activities via V158 and F158 Variant FcyRIIIa Receptors

Antibody-dependent cell-mediated cytotoxicity (ADCC) contributes to clinical efficacy of a broad range of therapeutic antibodies. FcγRIIIa polymorphism of individual cancer patients are correlated with clinical efficacy of some of these antibody drugs. Classic ADCC cytotoxicity assays rely on primary effector cells, which are highly heterogeneous and variable. To quantitatively measure antibody activity and evaluate the impact of FcγRIIIa polymorphism, we developed a pair of reporter-based ADCC assays using two engineered effector cell lines in Jurkat that stably express an NFAT-RE driven…

Regulation of cell apoptosis by insulin and IGF-I

An increase in the number of cells growing in cell culture is the result of two opposing effects: an increase in the number of cells that traverse the cell cycle and divide into two daughter cells (mitosis), or a decrease in the number of cells that die according to different modalities, the most prominent one being apoptosis (1), or both. Insulin and IGF-I are both capable of facilitating cell progression through the cell cycle, and of inhibiting apoptotic mechanisms. In…

Microbial Cultivation In Different Scales in the CELL-tainer® Rocking Single-use Bioreactor

Single-use bioreactors are applied in the biopharmaceutical industry for mammalian cell culture processes. The low gas-liquid mass transfer coefficient generally achieved in these systems does not allow for the compatable application of microbial processes in these bioreactor systems. The CELL-tainer® technology combines a vertical and horizontal movement in a rocking bioreactor concept, so that the turbulence and power input is intensified. In this system, mass transfer rates comparable to stirred tank reactors are achievable. Thus, volumetric oxygen transfer rates (kLa)…

A Systematic Strategy of Cell Culture Media and Feed Development

Development of a high-performance and robust fed-batch process for recombinant Chinese Hamster Ovary (CHO) cell lines presents challenges in light of the diverse nutritional requirements observed with different clonally derived cell lines as well as the varied production phases. To address these challenges, we will discuss an innovative strategy of media, feed, and process development that has been developed at FUJIFILM Diosynth Biotechnologies (FDB). The efficiency and effectiveness of this strategy have been well demonstrated in multiple mAb-producing CHO cell…

Next Generation Depth Filters for the Clarification of Flocculated Mammalian Cell Culture

Increasingly high cell density, high product titer cell cultures, containing mammalian cells are placing a larger burden on traditional downstream clarification and purification operations due to higher product and impurity levels. EMD Millipore has addressed these challenges by developing a proprietary single-use technology to improve primary and secondary direct depth filtration of flocculated high cell density mammalian cell cultures. The use of graded depth filters, combined with pre-treatment technologies, have achieved higher primary throughput capacities and lower flushing volumes as…

Liquid-Liquid Porometry of Viresolve® Pro Membrane

A liquid-liquid porometry (LLP) test was developed to characterize the pore size distribution of Viresolve® Pro membrane. Data generated by this test supports the understanding of the size-based virus removal mechanism of the filter. The LLP test also is used at-line during membrane casting. Process control based on LLP results improves membrane consistency.

Semi-automated, Single-use TFF Systems With an ‘Open Architecture’ Design Maximize Flexibility and Allow Supply Chain Security

Cross flow filtration used in bioprocesses is typically either microfiltration to clarify process streams or ultrafiltration to concentrate and diafilter the biopharmaceutical molecule. Single-use cross flow operations provide a number of advantages over traditional stainless steel systems. These include: • Flexibility during process development • Reduced capital investment • Avoidance of cross contamination and faster product change overs • Elimination of steaming and cleaning utilities Parker domnick hunter’s innovative SciFlex® TFF system utilizes propriety technology allowing the sterile processing of…