Course Code: PHRM 407
Course Title: Pharmaceutical Biotechnology
Department of Pharmacy
Nishat Tasnim Diba
Date of submission: 06-11-2018
PRODUCTION OF MONOCLONAL ANTIBODIES
Monoclonal Antibodies ((MABs) are specific type of antibodies which are made by their identical immune cells and that are the clones of a unique parent cell.
This Hybridoma Technology is a method for monoclonal antibody production. Identical antibodies as well as a large numbers of monoclonal antibodies (MABs) are produced by this process. Hybridomas are a type of cells which is engineered to produce a desired antibody in plenty amounts, and these products are the monoclonal antibodies. In 1975 two scientists, G. Kohler along with C. Milstein discovered Hybridoma technology and in 1984 they were awarded Noble prize for medicine and physiology.
• Hybridoma, is produced by the injection of a specific antigen into a immunized subject animal (e.g. mouse), procuring the plasma cells (antibody- producing cell) which are antigen specific. Plasma cells are from the mouse’s spleen and the subsequent fusion of this cell with a cancerous immune cell referred to as a malignant neoplasm (myeloma cell).
• Soon after the spleen cells are isolated from the mammal’s spleen, the myeloma cells and the B cells get fused.
• By this process hybrid cells are produced and it can be cloned to produce many identical daughter clones.
• Immune cell products are then secreted from those daughter cells. When these antibodies come from only one type of cell (the hybridoma cell) they are called monoclonal antibodies.
• This process can combine the qualities of the two different types of cells; the ability to grow continually, and to produce large amounts of pure antibody; that is the advantage of this process.
Figure: Production of MAbs by Hybridoma Technology
• HAT Selection
o HAT medium (Hypoxanthine Aminopterin Thymidine) is employed for preparation of monoclonal antibodies.
o Laboratory animals (mice) are 1st exposed to associate substance to that we tend to have associate interest in uninflected an antibody against.
o Once spleen cells are isolated from the vertebrate, the B cells are coalesced with immortalized malignant neoplasm cells that lack the HGPRT (hypoxanthine-guanine phosphoribosyl transferase) gene exploitation polythene glycol.
o Fused cells are incubated within the HAT (Hypoxanthine Aminopterin Thymidine) medium.
• Myeloma cells die, as they can’t turn out nucleotides by the de novo or salvage medium blocks the pathway that permits for nucleotide synthesis.
• Unfused B cells die as they need a brief lifetime.
• Solely the B cell-myeloma hybrids survive, since the HGPRT gene coming back from the B cells is purposeful.
• These cells turn out antibodies (a property of B cells) and are immortal (a property of myeloma cells).
APPLICATION OF MONOCLONAL ANTIBODIES
Now it is proven hat, Monoclonal antibodies are terribly helpful as diagnostic, imaging, and therapeutic reagents in clinical drugs.
Several monoclonal antibody diagnostic reagents currently obtainable are the right products for detecting various physiological, clinical or pathological conditions like pregnancy, diagnosis of various infective microorganisms.
By Immunoscintingraphy technique, radiolabeled Monoclonal Antibodies are used in diagnostic imaging for
• Myocardial Infarction (MI)
• Deep Vein Thrombosis (DVT)
• Cancer (Multiple Myeloma, Breast Carcinoma, Ovarian Carcinoma, Colorectal Carcinoma, Lung Carcinoma, Sarcoma etc.)
Clinical or in therapeutic application of monoclonal antibodies includes infectious disease, cardiovascular disease, cancer, auto-immune disease and also transplantation of bone marrow and organs by different forms of MAbs naming
• MAbs alone
• Radioisotope immunoconjugates
• Toxin and drug immunoconjugates