Downstream Processing

AbSolute® High Cap by Novasep: Revolutionary Protein A Media

Novasep has developed AbSolute® High Cap, an optimized silica-based protein A media for the capture of monoclonal and polyclonal antibodies. This protein A media is adapted to all fermentation volumes while offering the highest dynamic and static binding capacities (DBC10% ≥ 60 mg/mL, SBC ≥ 95 mg/mL) and, as a result, high productivities at all velocities. By decreasing the amount of resin required to downstream a feed (30-50% less), AbSolute® High Cap enhances the robustness of the protein A capture…

A New Mixed-Mode Resin for Large Scale Biomolecule Purification

Chromatographic resins with high capacities and selectivities differing from those seen with traditional hydrophobic interaction and ion exchange media are now in demand. Mixed-mode chromatography media is an alternative. Some mixed-mode resins combine both traditional hydrophobic interaction and ion exchange media. This poster introduces TOYOPEARL® MX-Trp-650M, a high-capacity weak cation mixed-mode resin for the purification of biomolecules. The polymethacrylic base bead was chemically modified with the amino acid tryptophan which combines a weak cationic group with a hydrophobic functional group.…

Purification of Antibody Fragments: A New Route for Capture

Antibody fragments have the potential to become the next important class of protein-based biotherapeutics after monoclonal antibodies (MAbs). The introduction of Capto™ L, with its protein L ligand, provides a possibility for an industrial platform purification approach for capture of this class of biomolecules. Capto L can bind a broad range of antibody fragments containing kappa light chains. Capto L is the latest member of chromatographic media (resins) in the toolbox consisting of several different products with affinity for the…

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…

Characterization of a Novel High-Capacity Weak Cation Exchange Resin

Ion exchange resins with increased selectivity and binding capacity are now in demand and it is imperative for chromatographers to have such resins in their repertoire. The Toyopearl GigaCap® family of resins was created to meet these demands. This poster will focus on Toyopearl GigaCap CM-650M resin as a novel high-capacity weak cation exchange resin for the purification of biomolecules. A polymethacrylic base bead, Toyopearl HW-65, was chemically modified with carboxymethyl groups in the 1000Å pores of the bead which…

Evaluation of Ultra Performance Size Exclusion Chromatography for the Analysis of Proteins and Higher Order Aggregates

Complete characterization and analysis of biopharmaceuticals includes size exclusion chromatography (SEC) to measure protein aggregates and other size variants. Current silica-based HPLC-SEC methods can be time-consuming and unreliable. New advances in packing materials and instrumentation have allowed faster and more reproducible separations to be achieved. Improvements in resolution, sensitivity and throughput will be demonstrated for sub-2 μm SEC packing materials as compared to traditional silica-based columns. The effect of pore size and particle size for the analysis of aggregates and…

When downstream process is able to withstand upstream changes! An example of clarification sequence optimization in viral vaccine production

While improving cell growth and viral productivity in USP, DSP may become the production bottleneck if not able to withstand changes. Such a case appears when optimizing cell culture media during the development of a viral vaccine candidate. Thirty percent expression increased was balanced by twenty percent yield loss and filters clogging during the clarification step. As a consequence, clarification sequence was not scalable anymore for the further manufacturing. The poster describes sanofi pasteur approach to overcome this issue and…

Lipid Removal by Depth Filters in Plasma Fractionation

We herein describe a 3M Purification Inc. depth filter media – Zeta PlusTM DELI that exhibits selective adsorptive properties for plasma lipids. Lipids plug chromatographic columns and filters during plasma fractionation steps and cause solution instability for the final product. Several parameters which could affect the lipid removal efficiency on Zeta Plus DELI have been investigated: prefiltration, contact time, ionic strength, pH, and temperature. The maximum percentage of total lipids eliminated, in optimal operating conditions, was 68%. This method has…

Salt-tolerant cation exchanger for direct capture of proteins

Ion-exchange chromatography is widely used for the purification of biotherapeutic proteins. Usually, cation exchangers are applied in the primary capture step of proteins with alkaline pI. Conventional ion-exchange chromatography has the advantage of high-binding capacity, but it requires low salt conditions for protein binding and consequently adjustment of conductivity of clarified culture supernatant. An alternative method to maintain some binding capacity at higher conductivity levels is working at low pH condi¬tions notwithstanding that some proteins are sensitive to acid treatment.…

Protein A Cellulose, a new mAb purification platform

Protein A chromatography is widely used as capture step in monoclonal antibody purification processes. Many types of chromatographic media are commercially available for this application however mainly Agarose and Porous glass based products are considered as standard mAb purification platforms due to high dBC and high operational flow rates. In fact process optimization of Protein A step is mainly aimed to have higher capacity and lower elution volumes in shorter process time. In this context Kaneka has been investigating highly…