Posters

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…

Continuous Bioprocessing: A CMO’s Perspective

The maturation of many process industries is marked by transition from batch to continuous processing. Often this move to con-tinuous processing determines the winners in the industry. Will the bioprocess industry go the same way? The analogy is often drawn to the chemical industry where continuous processing is widely applied. Although available for many decades continuous bioprocessing is not yet a mainstay of the industry. Currently there is a ground swell of feeling within the bioprocess industry that the time…

Scalable Method for Packing Chromatography Columns

Currently the bio-pharmaceutical industry is heading towards cost cutting, time saving and single use manufacturing approach. This has opened up a whole new market for disposable technologies. Pre-packed disposable chromatography columns are a best fit for preclinical and clinical stage Monoclonal Antibody (mAb) production and vaccine manufacturing processes which require quick turnaround times and aggressive validation respectively. This work describes development of pre-packed and ready to use chromatography columns to meet this need in the industry. A robust packing method…

Expansion of T-cells using the Xuri Cell Expansion System W25 and WAVE Bioreactor 2/10 System

Immunotherapeutics include drugs and biologics that render therapeutic benefit by harnessing the power of the immune system. The promise of immune-mediated therapies is to target specificity with a consequent reduction in off-therapeutic effects. Immunotherapeutic products can be classified broadly into (1) active immunotherapy (therapeutic vaccines), (2) adoptive cellular immunotherapy (transfer of immune cells, genetically modified T-cells or precursor cells) or (3) passive immunotherapy (antibody or receptor ligand administration). Recent scientific advances have led to clinical trials of both active and…

The Importance of Thresholding in Imaging Analysis of Protein Aggregates

Dynamic imaging particle analysis (DIPA) shows much greater sensitivity to transparent particles, such as protein aggregates, than light obscuration can. While not yet fully accepted in industry compendia, DIPA is being used increasingly in the formulation process for characterization of sub-visible particulates in biologics. DIPA measures particle size and shape by first creating a binary image based upon a defined threshold from the background value for each pixel in the image. The resulting binary image is used for all particle…

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…