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…

Platform Purification of a Domain Antibody

Monoclonal antibodies (MAbs) are very successful for treatment of several different cancers and tumors. However, the low tissue penetration has led to the development of smaller sized biopharmaceuticals (Ab fragments) such as Fabs, single chain Fv (scFv) and domain antibodies (Dabs). These molecules lack the Fc part of the antibody making a platform purification approach using Protein A impossible. However, with the introduction of the Protein L based affinity chromatography media (Capto™ L) new possibilities are introduced for capture of…

Aggregate Removal from Monoclonal Antibody with Nuvia™ HR-S Media

Nuvia HR-S media is a new cation exchanger designed for high resolution of closely related product impurities such as aggregates. It delivers excellent resolution, with a final aggregate content of <0.3%, and high recovery of >80% from a heterogeneous feed of monoclonal antibody aggregates and monomer. Aggregate content and recovery in the eluate were shown to be functions of the target conductivity measured at the end of collection.

Enhancing Efficiency and Economics in Process Development and Quality Control of Biotherapeutics

Analytical techniques that measure protein quantity and quality are used in nearly all stages of research, process development, quality control and manufacturing of biotherapeutics. UV spectroscopy, ELISA and HPLC have been in use for decades for protein quantitation in physiological and process samples, and continue to be the workhorses despite their many limitations. Biopharmaceutical companies have enthusiastically adopted Pall ForteBio’s Octet® systems due to their high throughput capabilities, decreased sample preparation requirements, and low cost of operations. The Octet systems…

A Case Study: 3-step Process for Efficient mAb Purification

This study showcases a portfolio of commercially available biopharmaceutical chromatography resins designed for the efficient purification of monoclonal antibodies. A 3-step purification process has been implemented which showed effective removal of the main contaminants, low ligand leakage, and high yields over the entire process. Eshmuno® A affinity chromatography resin was evaluated as the first step in the process. The Protein A elution pool was further purified using cation exchange chromatography. Two cation exchange resins with different selectivities were compared. The…

Progress On A Fully Disposable Downstream Platform: A Simple, Risk-Free, Plug-in Solution To Solve The DSP Bottleneck

Natrix HD Membrane technology features a polymeric hydrogel formed within a flexible porous support matrix. The support matrix provides mechanical strength, while the hydrogel properties determine the separation chemistry of the product. An advantage of the Natrix chemistry is the ability to place virtually any functional group chemistry throughout the hydrogel polymer. Natrix HD membrane technology combines the superior binding capacity of conventional resinbased columns with the high throughput of membranes in a single-use format that eliminates costly packing, cleaning,…