In an effort to improve the antibody manufacturing platform, newer Cation Exchange Resins were evaluated. Dynamic binding studies were conducted using 2 different monoclonal antibodies at a stable pH and varying conductivities across nine resins to determine the differences in binding capacity. DBC Studies were performed at a pH of 5 and conductivities of 5, 10 and 15 mS/cm for the nine cation exchange resins with breakthrough criteria set at 10%. While most of the resins provided excellent binding capacity…

BAC BV – The Affinity Experts – develops novel & cost-effective solutions for affinity purification challenges. Affinity chromatography is one of the simplest and most effective methods for purifying protein therapeutics, offering reduced process steps and higher yields than non-affinity methods can provide. For standard MAb purification Protein A is a well established affinity ligand which clearly has demonstrated the benefits of a highly selective primary capture step. However, for non antibody based therapeutics and novel antibody formats lacking a…

Ceramic hydroxyapatite (CHTâ„¢) is used for protein purification owing to its superior removal of all process-stream impurities (aggregates, HCP, viruses, endotoxin, protein A, DNA, etc.). At commercial scale CHT performance can be impacted due to the release of protons during elution. This proton release lowers mobile phase pH which may impact the robustness of CHT particles. We have developed a novel, patent-pending technology called the surface neutralization system (SNS), which desorbs protons from the surface of CHT prior to elution.…

Clarification and capture challenges are inherent in a new cell culture process that extends the exponential growth phase of mammalian cell cultures, resulting in high cell densities and product titers. Here, we show that second generation Expanded Bed Adsorption (EBA) technology is ideal for processing such high titer, high cell density harvests. In these culture harvests, viable cell densities of > 100 million cells/mL and product titers > 10 g/L are observed routinely, using standard media and cell lines. Several…

APT1604 is a base-stable 600 Da peptidometic ligand coupled to a 6% cross-linked agarose matrix. APT1604 binds to Fc-fragment of monoclonal antibodies (mAb) and various immunoglobulin (IgG) subclasses. The interaction between protein and ligand has been suggested to result from mixed mode ionic – hydrophobic interactions, albeit with high selectivity for immunoglobulins. The main application area for APT1604 is purification of mAb from mammalian cell harvest, but it can also be used for purification of IgG from other sources. The…

In this work we present results with a novel high capacity Protein A medium prototype based on high flow agarose. The medium prototype was optimized for higher dynamic binding capacity (DBC) at long residence time. A 20–50% higher DBC was obtained compared to MabSelect SuReâ„¢. DBC was independent of MAb concentration in the range 1–10 g/L. Optimization of elution conditions (pH and flow rate) was performed by design of experiments. A lifetime study for 100 purification cycles, performed with a…

Reliable analytical tools are valuable for economical process development, production, and batch release of vaccines. The current method recommended by the European Pharmacopoeia and WHO for determination of influenza virus titer ¬ single radial immunodiffusion assay (SRID) is straightforward and easy, but leaves plenty of room for improvement when it comes to assay variability and speed of analysis. In addition to having low sensitivity and precision, the method is both time-consuming and quite labor-intensive. In this study a Biacoreâ„¢ assay…

With increasing use of single-use, disposable fluid processing systems, concerns of mixing arise in processes such as protein concentration, clarification and diafiltration. For some operating conditions of flow rate, solution viscosity or flow path configurations, protein solutions may become stratified in vertically-oriented hanging bags. This can cause decreased efficiency in cross flow applications. In this study, we modeled a stratified solution and tested the influence of these various operating conditions using ReadyCircuit* 2-D pillow bags. Even with viscous materials, flow…

Much attention has been given to ion exchange (IEX) media as a means to improve productivity as a result of increasing demand for higher efficiency on the downstream process. Until recently, strain optimisation for high productivity and upstream purification were the bottlenecks for most bio-processes. However, with the progress made in recent years, titers in fermentation processes have increased significantly. Obviously, this increased volumetric productivity will help reducing the cost of goods, but it also has an impact on the…