Agenda

11:15 – 11:45

Detection of Low Affinity Anti-Drug Antibodies and Improved Drug Tolerance in Immunogenicity Testing by Octet^® Biolayer Interferometry
Jian Li, Research Scientist, Centocor

Abstract
Anti-drug antibody (ADA) immunogenicity assessment is a critical component in demonstrating the safety and efficacy profile of a therapeutic biological drug.  Double antigen bridging immunoassays are widely used to measure ADA, however, these methods can generate false negative results due to low sensitivity for the detection of low affinity ADAs and interference from high concentrations of biological drug in samples.  The goal of this study was to develop a method that could detect ADA to a human therapeutic antibody in the presence of drug within the sample.  Using Octet QK and mock ADA samples we demonstrated that this method offered advantages over ELISA and ECLIA methods.  We then investigated the capability of Octet RED to detect ADA in a pre-clinical toxicokinetic and tolerability study of CNTO X mAb and compared it to ADA detection by ECLIA, leading us to believe that the Octet biolayer interferometry platform offers much promise for the development of drug tolerant ADA immunoassays.   Experimental results of our study will be presented.

11:45 – 12:15

Ligand Binding Assays on the Octet Platform for Bioprocess Contaminants: Residual Protein A and HCP
Sae Choo, Principal Scientist, ForteBio

Abstract
ForteBio’s Octet Dip and Read™ assay platform provides fast, accurate and easy-to-use assay solutions for many bioproduction applications. Direct binding assays that detect proteins without the need for secondary reagents are complemented by multi-step assay methods, affording a large dynamic range from picograms/mL to milligrams/mL. Octet systems are currently utilized in various stages of biopharmaceutical manufacturing to replace HPLC and ELISA methods which suffer from long analysis times, lack of specificity, labor-intensive protocols and imprecision. This talk will highlight Octet assays to detect residual host cell protein contamination in CHO cells and residual protein A contamination in antibody preparations.

12:15

Lunch

12:20 – 12:45

Using Biolayer Interferometry to Identify Wnt/LRP6 Signaling Complex Inhibitors and Decipher Their Mechanism of Action
Eric Bourhis, Scientist, Genentech

Abstract
Wnt/β-catenin signaling is initiated at the cell surface by association of secreted Wnt with its receptors Frizzled (Fz) and LDL-receptor-related protein 5/6 (LRP5/6). Study of these molecular interactions has been a significant technical challenge because the proteins have been inaccessible in sufficient purity and quantity. Here, we describe insect cell expression and purification of soluble human LRP6 extracellular domain and its use in screening Wnt/β-catenin signaling inhibitors. Direct binding assays using the Octet RED label-free assay system revealed the existence of multiple, independent Wnt and Dkk1 binding sites on the LRP6 co-receptor, suggesting new possibilities for the architecture of Wnt signaling complexes and a model for broad-spectrum inhibition of Wnt/β-catenin signaling by Dkk1. Additionally, we show that we can use this assay to screen antibodies blocking LRP6 function and predict their cellular activity based on the subset of Wnt they inhibit.

12:45 – 1:15

Biopharmaceutical Production Using Pfenex Expression Technology™ and High-Throughput Biolayer Interferometry
Jeff Allen, Director, Analytical Biochemistry, Pfenex

Abstract
The Pseudomonas-based Pfenex Expression Technology has proven to be a robust and cost effective platform for the production of numerous classes of therapeutic proteins, including various types of antibody derivatives.  High cell densities in the fermentor along with high specific protein yields result in high target protein titers.   Very high cell densities are also achievable even in small scale growth (96-well plates).  An extensive tool box of unique host strains has also been developed.  These strains have phenotypes selected to impact the amount of target protein produced and its solubility and activity.  Coupled to an automated sample preparation platform, parallel through-put biolayer interferometry (BLI) allows hundreds of different expression strategies and host cell combinations to be rapidly tested for both quantity and functionality. 

1:15 – 1:30

Speaker Panel Discussion