Posters

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…

Viral Vector Production in the Integrity® iCELLis® Disposable Fixed-bed Bioreactor from Bench-scale to Industrial Scale

Recombinant viruses (e.g. lentivirus and adeno-associated-virus) can be used as human gene therapy vectors. They are mainly produced in adherent cell cultures (e.g. HEK293T, A549, VERO) in Roller Bottles or multiple-tray-stacks using either transient transfection or infection strategies. Therefore, the Integrity® iCELLis® (ATMI LifeSciences) line of bioreactors offers a new production alternative with stronger process controls and ease of scale-up.   The iCELLis bioreactor is designed for adherent cell culture applications. Cells grow on microfibers carriers packed in a fixed-bed…

Is the Gold Standard Tarnished? Weaknesses in the Standard Method Compromise Your RMM Validation

Introduction of a new Rapid Microbial Method (RMM) requires a full validation. The purpose of the validation is to prove that equivalent or better results are obtained using the new method when compared to the compendia. One assumes that the result obtained with the compendial method is absolute and is the “Gold Standard” by which to compare the new test. In reality, the compendial method exhibits a number of weaknesses that compromise the integrity of the validation. The standard requires…

Cookie Cutter Proteolysis: Achieving Reproducible, Efficient Digestions for Proteomic Workflows

Cookie Cutter Proteolysis: Achieving Reproducible, Efficient Digestions for Proteomic Workflows   Protein sample preparation workflows for mass spectrometric analysis that involve proteolysis are often labor-intensive, time consuming and user dependent. These workflows often involve digestion, solid phase extraction, drying, and re-suspension prior to reversed phase separation into the mass spectrometer. The introduction of variability at many of these steps hinders discovery initiatives as well as the ability to convert these discoveries into viable assays.   Recently, an automated protein digestion…

Maximizing Protein Expression in CHO Suspension Cells Through Transient Transfection

Transient transfection in mammalian cell lines provides an avenue for researchers to bridge the development bottleneck and shorten the time to usable protein. The method also maintains post translational modifications crucial for biotherapeutic function. Chinese Hamster Ovary (CHO) suspension cells are especially suited for high yield production of recombinant proteins, despite being refractory to commonly used transfection methods (e.g. 25kDa linear PEI). Mirus Bio has developed a more effective alternative, the TransIT-PRO® Transfection Kit. Transfection efficiencies are affected by many…

T Cells Expanded in the WAVE Bioreactor™ 2/10 System Maintain a Healthy Phenotype

T cell immunotherapy often requires the expansion of a small select starting population in vitro. To achieve therapeutic doses, this population is required to undergo multiple and rapid rounds of replication. Rapid T cell expansion raises the possibility of inducing senescence or an aged phenotype, both of which are detrimental to the recipient patient. The WAVE BioreactorTM System is often used for the final expansion phase before patient infusion and we have analysed the aging characteristics of T cells that…

Transfer of Hepatic Progenitor Stem Cell Culture Process from Multitray Stacks to the Integrity® Xpansion™ Multiplate Bioreactor

Scale-up a stem cell process may be challenging: small variations in physicochemical parameters (surface characteristics, pH and dissolved oxygen) can heavily impact stem cell growth and behavior. The Integrity® Xpansion™ multiplate bioreactors have been designed to enable an easy transfer from multiple-tray stacks process by offering the same cell growth environment: stacked hydrophylized polystyrene plates in a compact and closed system (from 10 to 200 plates per bioreactor equivalent respectively to 6120cm² and 122400cm²). As there is no headspace between…

Comparison of Commercial Kits for Extraction of Residual Host Cell DNA

Expression of therapeutic proteins in cultured cells is a cost effective method for production of commercial quantities of a drug substance. However, the manufacturing and purification process of these products leaves the potential for DNA contamination from the host cells. Due to the theoretical potential for the transfer of oncogenes from the host cell, the WHO has set a residual host cell DNA limit of 10ng/dose in biologic therapeutics. Regulatory agencies have set allowable limits between 100pg/dose and 10ng/dose depending…

New Core Bead Chromatography Medium Enables Group Separation and High Productivity Purification of Large Molecules

When large biomolecules such as IgM or influenza virus are purified, group separation with gel filtration is the most commonly used technique. Advantages of using gel filtration include the high purity and the high recoveries that can be achieved, while a drawback is the relatively low productivity. Capto™ Core 700 is a chromatography medium (resin) from GE Healthcare Life Sciences designed with core bead technology. The core bead technology enables double functionality for Capto Core 700 – size exclusion and…

Improving Cell Culture Productivity with a SEGFLOW® Automated On-line Sampling and Feed System

A SEGFLOW® automated on-line sampling and feed system (SEGFLOW) was evaluated for its impact on Chinese Hamster Ovary (CHO) cell culture process performance in bench-scale stirred tank bioreactors. The SEGFLOW was integrated with a YSI® Biochemistry Analyzer and a nutrient feed pump allowing real-time glucose analysis and automated glucose feeding to be performed for the test batch. In contrast, a batch using daily manual sampling and bolus feed protocols, was performed as a control. For the SEGFLOW culture, media glucose…