Chromatography is a useful purification method for large biomolecules and virus manufacturing and it is easily scalable to large production volumes. Convective Interaction Media (CIM) monolithic columns constitute of large flow-through channels and consequently have high surface accessibility of binding sites. Preferences of CIM monolithic columns are flow independent performance, resulting in fast separation, concentration, purification, impurities removal, and analytics of biopharmaceuticals.
The aim of the study was to develop Influenza virus purification platform, which can be used for several virus strains. The main objective was to develop a process with as little as possible of intermediate steps, especially omitting Tangential Flow Filtration (TFF) or other sample pre-treatments with high host-cell DNA and protein removal, as well as to achieve high binding capacity of the Influenza virus per mL of monolithic support.
CIMac™ r-Protein A Analytical Column is a short bed, high performance monolithic column. It is intended for fast, efficient, and reproducible qualitative and quantitative analyses of Immunoglobulin G (IgG) and suitable for use with HPLC and UPLC systems. Quantification of IgG is possible between 0.2 μg and 20 μg. Its small volume and short column length allow operation at high volumetric flow rates (up to 3 mL/min). The information about product quantity and purity is thus generated in just 1 minute! The column has an innovative symmetric design for bi-directional flow contributing to longer lifetime.
Hydrazide-activated (HDZ) columns were proven to be a product of choice for making the most effective immunoaffinity columns. They take advantage of a special hydrazide linkage that binds antibodies through the carbohydrate residues on their Fc regions. This leaves the antigen-binding domains fully accessible to enable the most effective capture of desired target.
Hydrazide-activated (HDZ) columns were proven to be a product of choice for making the most effective immunoaffinity columns. They take advantage of a special hydrazide linkage that binds antibodies through the carbohydrate residues on their Fc regions. This leaves the antigen-binding domains fully accessible to enable the most effective capture of desired target (Figure bellow).
CIMac™ HDZ monoliths make HDZ-immobilized antibody columns even more effective. Because of their large channel size and the efficiency of convective mass transport, they eliminate the long loading residence times that are required for affinity chromatography on porous particle columns. Flow rates of 5–10 column volumes per minute allow complete purifications in a few minutes, even when the source material contains a low concentration of antigen. The same performance is achieved whether a small peptide or a large bio-assemblage like a virus particle or extracellular vesicle is isolated. The combination of HDZ monoliths and the immobilization protocol offers a strong tool for fast antigen isolation from complex biological sample (plasma, lysate, etc.) and consequently sensitive antigen quantification. An example of CIMac™ HDZ application is a purification of fibrinogen from human plasma.
There are many cases, where a single protein needs to be purified from a complex sample. Such proteins manifest themselves as impurities, which can affect further analysis, either by causing specific equipment malfunction or lower yield in the products. In other cases the specific protein is our molecule of interest, for example in glycomics analysis. In both cases high specificity for proteins, reproducibility and reliability is necessary. We have developed a model immunoaffinity column and 96-well plate based on an anti-fibrinogen monoclonal antibody, covalently immobilized onto CIMac™ HDZ analytical chromatographic monolith.
Yes. Make sure that either the inlet or the outlet of the column is open to prevent excess pressure buildup. Autoclaving monolithic columns for aseptic use is recommended. Prior to disposal, monoliths can also be autoclaved. For specifics, please consult firstname.lastname@example.org.