Future of Biopharmaceutical market for Therapeutic Monoclonal Antibodies looks very bright. Since the award of a Nobel Prize for the technology that enabled the preparation of monoclonal antibodies in 1984, their utility has expanded far beyond their scientific use; they have become highly valuable therapeutic agents. Monoclonal Antibodies directed against cancer and autoimmune diseases are burgeoning and this is a huge growth area due to the success of drugs already on the market. Thus, the use of monoclonal antibodies is expected to grow exponentially in days to come. Generating annual sales of almost $80bn per year today, monoclonal antibodies continue to play a vital role in the biopharmaceutical development and represent 42% of the total pharmaceutical market.
Monoclonal Antibody(Immunoglobulin G, IgG2a, mAb) molecule, chemical structureThis is primarily driven by the oncology and autoimmune disease treatments. A series of Animal Room Facilities, including Specific Pathogen Free Barrier facility, Transgenic Animal Facility, will be used for Monoclonal Antibody Vaccine production, infectious disease research and Gene Knockout Studies.
The overall goal of our mAbs program is to develop a reliable and reproducible process that enables production of a mAbs product that is safe to administer to humans. A critical aspect of the development of any mAbs is the selection of a suitable genetically modified, engineered cell line capable of producing sufficient quantities of the product. Many emerging or established technologies are available to improve the speed of cell line development or the specific productivity levels that can be achieved. After the production cell line is established, a cell culture, or upstream, process that enables production at the desired scale will be developed. The upstream process enables expanding the initial cells up to a sufficient biomass to inoculate the bioreactor, growing the cells to maximum density and viability, and supporting maximum productivity.
Upstream process development includes systematic optimization of the growth and production culture medium and the bioreactor conditions to produce the maximum amount of active product of desired quality with the minimum amount of resource consumption. Some refinements of the cell culture process may be required at the production scale. The need to obtain data at the production scale prior to finalizing all aspects of the process make it essential to conduct non-cGMP pilot, or engineering, runs at scale prior to producing clinical material under full cGMP conditions.
Tumor (Myeloma) cells that can replicate endlessly are fused with mammalian cells that produce an antibody. The result of this cell fusion is a "hybridoma", which will continually produce antibodies. These antibodies are called monoclonal (clones of one molecule, hence identical) because they come from only one type of cell, the hybridoma cell. The next stage is multiplication of the cells by cell culture. The multiplication is carried out in bioreactors and the cell mass is purified to get the MAbs. Antibody manufacturing starts with the embedding of antibody production genes in the host cells. The most popular types of host cell are CHO cells, NSO cells, and SPII / O cells (the last two both being mouse myeloma cells). The cloned cells containing the antibody-producing genes are then screened, and the cells that produce the largest volume of antibody are used to establish the master cell bank (MCB).
The cell cultivation stage involves first establishing a master cell bank (MCB), and then discovering the optimal composition of the culture medium. The cells obtained from the MCB are cultivated in a flask or other small container. Plastic bags are then used for cultivation on a scale of several tens of liters, before moving up to disposable bags/ Stainless steel bioreactors with a capacity of several thousand liters. The cell cultivation stage usually takes 2-3 weeks, depending upon the cells involved and the production scale. The culture medium composition has a major impact on productivity. Determining the optimal composition of the culture medium appears to be a very time-consuming job, and is thus the area that requires the most technological collaboration. The next process after incubation is the removal of cell material. Large impurities such as cell membranes are removed using a disc stack centrifuge, into which liquid can be fed continuously, and then the solution is passed through a depth filter (a special filter that catches materials on the inside of a membrane). Lastly, large contaminants are removed from the antibodies using micro filtration with a pore size of 1 micron or smaller. The filtered solution then passes on to the purification process. The process is similar to processes used for animal cells as well as E. Coli and other bacteria.
The final process in antibody manufacturing is purification, which removes substances other than antibodies with particles sized smaller than 1 micron, including proteins and DNA fragments. Normally, the first step would be to use precision elution to separate the antibodies in a chromatography column with protein A binding (Protein/Cation-Exchange chromatography). The second step is to achieve the quality required for the final products using low-pH viral inactivation, the separation of protein and DNA other than the antibodies using anion exchange chromatography. Chromatographic purification is followed by virus filtration, ultra filtration and sterile filtration.