BioProcessing 2019

Ambrx’s mammalian expression platform (EuCODE™) enables non-native amino acids (nnAAs) through an expanded genetic code to both generate novel bio-therapeutics and to optimize the performance of antibody drug conjugate (ADC), therapeutic proteins, monoclonal antibodies (mAbs), and, bi- and multi-specific medicines. While the ability to control the defined Drug-to-Antibody Ratio (DAR) and payload site can provide an advantage to an ADC, the site-specific incorporation of the nnAAs into the antibody heavy chain introduces a unique challenge for antibody production.

To enable higher performance benchmarks in time and resources for process development with stringent product quality requirements, a proprietary cell culture platform is being developed and demonstrated fast-track development of high-quality, high-titer processes for producing recombinant proteins from CHO cells. 

We successfully generated a CHO-K1 cell line, stably expressing engineered amber suppressor tRNA and its cognate tRNA synthetase specific for nnAA para-acetyl phenylalanine (pAF), to achieve high production of monoclonal antibodies (mAbs) containing nnAAs. The stable cell lines were further evolved using CRISPR/Cas9 genome editing technology to sequentially knock out selected genes in glutamine synthesis, and, apoptosis pathways to improve selection efficiency and prevent loss of viable cell mass in production cultures, respectively.  Inhibition of apoptosis pathway leads to dramatic increase in viable cell mass and results in extended production time and increased productivity. 

In this presentation, we will discuss overcoming the challenges in cell culture platform development including cell line engineering, systematic DoE-based approaches on optimal chemically defined media and cell culture processes, and, strategies for scale up to clinical and commercial scales.

Learning Objectives: 

1. Learn what is cell culture platform and how to development your own cell culture platform
2. Learn the strategy to develop your own chemically-defined media

The biologics development process is fraught with risks from pre-clinical studies through clinical evaluation. Prominent among these risks are changes in the critical quality attributes of the biologic during the drug development process.  Here, we present data on the use of the ExpiCHO expression system for high titer transient expression of candidate therapeutics through creation of stable cell lines for large scale manufacturing.  By using the same cell line from transient to stable production, the risk of product quality drift can be reduced during the transition from pre-clinical to clinical development.

Multi-mode chromatography has come to prominence in recent years due to the general utility of multi-mode ligands for product capture and polishing. In particular, their application in flow-through polishing has significantly improved process economy. However, significant optimisation of binding and elution conditions is often required to achieve the desired level of purity.

To overcome the deficiencies of individual multi-mode ligands, we have developed a diverse library of Multi-Mode Mimetic Ligands™  incorporating both ionic and hydrophobic groups, which offer significant flexibility in use. The Multi-Mode Mimetic Ligand™ Library is organised into a 12 x 8 array micro-column block and comprises a diverse array of ligands with a broad spread of charge, hydrophobic and hydrophilic functionalities. The adsorbents are produced using an agarose support matrix and incorporate triazine attachment chemistry for ease of synthesis and superior stability, Screening can be performed in high-throughput mode using robotic liquid handling systems or by use of conventional hand-held multi-channel pipettes. This approach allows for rapid screening, and identification of potential ligands for a specific target. Furthermore, adjustment of the starting protein fraction to suit individual ligands is not required as the diversity of the library enables the identification of binding ligands which are best-suited to the existing conditions and impurities.

This approach has been evaluated in a variety of applications, including development of ligands for the removal of specific contaminants in a flow through mode.

The Strep-Tactin®XT:Twin-Strep-tag®-purification system enables protein purification at high yields and purity under physiological conditions. Providing the highest binding affinity among all affinity tag systems, the technology fulfills the demands of mammalian expression systems (e.g. Expi) and is well suited for downstream applications like SPR or the switchSENSE® technology.

Learning Objectives: 

1. Purification of Twin-Strep-tag® fusion proteins from Expi293TM and ExpiCHOTM supernatants using Strep-Tactin®XT
2. Learn how the Twin-Strep-tag®/Strep-Tactin®XT system can simplify kinetic studies using SPR or dynamic Biosensors switchSENSE® technology 



 

As the pace of biotherapeutic approvals increases, so does the pressure on makers of new biologics to complete their development processes faster and more efficiently. The success of the drug development process is directly proportional to the utilization of accurate, sensitive and fast methods for selection, identification, quantitation, purity and other analyses of the drug components and the final drug product. The common adage, “time is money” has never been more applicable than to today’s biopharmaceutical industry and its focus on improving bioprocessing technologies to reduce costs and increase efficiencies. By offering solutions that address the key stages of bioprocessing—from cell line development to process optimization and formulation to final product characterization—Bio-Techne and its family of brands streamlines measurement and quantification of the complex interactions between a drug product and its manufacturing process. Let us show you how our technologies and systems increase productivity and reveal new insights to accelerate the time to market for new biologics. 

By attending this webinar, attendees will gain more insight into the current state of the bioprocessing industry and learn how they can overcome difficult bioprocessing challenges with products and services from Bio-techne and its family of brands, R&D Systems, ProteinSimple, Novus, Tocris and ACD.

Learning Objectives:

1. Gain more insight into the current state of the bioprocessing industry 
2. Learn how researchers can overcome even the most difficult bioprocessing challenges with Bio-Techne’s products and services