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Abstract Monoclonal antibodies are one of the most important products of biotechnology and laboratories and companies all over the world are pursuing their large-scale production. However getting higher yields of monoclonal antibody (MAb) is a problem in Hybridoma Technology, which has two major bottlenecks: • Poor yield of hybridized cells • Low cellular productivity of MAb in culture. The behavior of hybridoma in culture is influenced by sophisticated cellular metabolic activities and various interactive environmental factors and that the understanding and modeling of the way hybridoma grow in the bioreactor should enable optimization of bioreactor operating conditions to achieve maximum monoclonal antibody formation However, due to the lack of the incomplete knowledge of hybridoma cultivation process, there exist many limitations and challenges to the advent of applications of process control and optimization in this field. There are three ways of obtaining high MAb yield in vitro: • Large scale culture • High density culture • Enhancing individual cellular productivity in culture Currently, the focus is on the correct synergistic combination of fortified nutrient media, bioreactor design and mode of operation. Maximization of cell culture longevity, maintenance of high specific antibody secretion rates, nutrient supplementation, waste product minimization and control of environmental conditions are important parameters for improvement of large scale production of MAb. However, the production of small quantities of monoclonal antibodies and recombinant proteins was carried out using a new low cost production system, the Super Spinner. The hybridoma technique allows monoclonal antibodies of a single specificity to be obtained in virtually unlimited amounts. In principle, monoclonal antibodies can be made against any cell macromolecule and so can be used to locate and purify the molecule and, in some cases, to analyze its function. These MAb’s can be of therapeutic or diagnostic values either in the medical fields (e.g. diagnostic of viruses, malignances, laboratory titration of antigens, detection of fibrosis, cirrhosis, metastasis of tumors infectious diseases and as magic bullet etc.). Or in environmental fields where, the production of MAb’s can be directed to detect tracers of antigens present in environmental e.g. pesticides pollution to water, vegetables, fruits or animals blood and tissues. Another application is to detect air pollution e.g. air filtrated on reference nitro-cellulose filters can be titrated using MAb’s and detects the dose of air pollution. This technology can be applied also to identify any environmental insult from other sources whatever industrial non intended or intended induced. Herein we report an efficient and economical protocol for hybridoma cell cultivation to produce monoclonal antibodies in tissue culture flasks; small glass cylinders (super spinner) and bioreactor. This leads to the production and purification--in order of grams--of one MAb intended for human therapeutic use or as diagnostic tools. This protocol proved to be simple, reproducible and cost effective by using of serumfree medium culture and amino acids optimization for the production of MAb. |