subject: Purification Of Monoclonal Antibodies [print this page] There are several uses for antibodies and antibody fragments in therapeutic and diagnostic applications and research. This is possible because of the number of ways in which the antibody and antigen are capable of interacting and the ability to control this interaction. Recombinant technology allows a limitless number of combinations between immunoglobulins, tags and certain proteins, making it easy to manage these molecules flexibly.
Monoclonal antibodies or mAbsare homogenous immunoglobulins synthesized by hybridoma cells. They find their use in the fields of immunodiagnostics and immunotherapy as they can be specific for a particular antigenic determinant of the target molecule.
The need for monoclonal antibody sequencing calls for the development of low cost production, with a special focus on experimenting with new culture media without using serum.
The role of serum
The goal of working with hybridoma cells is the purification and categorization of monoclonal antibodies. Avoiding the use of serum helps the purification process.
Serum guards the cell surface with enzymes, protecting it from damage. Serum also tends to change the physiological and physicochemical attributes of the cultures ambiance. Although serum lets the cell grow and stay viable, there are certain elements in it that can change its composition, depending on the source of the cell. This makes it necessary to routinely test the batches. By adding serum to the culture, there are possibilities of changes in the culture medium, resulting in side effects in the clinical application and also higher costs.
There are many methods of preparing serum-free media. One way is by including certain elements to the basal nutrient media which provide buffered low molecular weight compounds. This can be used in purification of monoclonal antibodies.
Protein production through recombinant DNA technology is a growing industry. The cost of purification is high and therefore, closely monitored. This is one of the challenges in this process and efforts are under way to develop viable processes to maintain product purity and viral clearance by incorporating certain processes. An increase in the titers for monoclonal antibody feed streams has necessitated the development of newer products and techniques or at least making the existing ones flexible enough to meet the challenges. Identifying monoclonal antibody sequence is the starting point of antibody engineering.
Therapeutic use of antibody sequencing
Purification of proteins for therapeutic uses needs the right technology to achieve the high level of purity that biopharmaceuticals insists on. The magnitude of the process calls for attention to technology design and implementation and a close control over the variables in the process since proteins degrade easily in extreme pH and temperature or when they come in contact with solvents. The cost of monoclonal antibody sequencing can be monitored by faster operations. Some monoclonal antibodies are required in large quantities annually and to meet this demand, an integrated series of purification stages must be designed. This will not only help increase the yield, but also make the product molecule specific.
Strategies to improve the efficiency of the process
A number of innovative strategies are being experimented with. These use disposable equipment, new filters and absorbents containing substrates that are lower priced but are more useful in lipid binding. The biggest challenge faced in monoclonal antibody sequencing is choosing the right process from the number of technologies available.
