![]() ![]() Any discomfort, stress and pain should be minimized when this method is being used, and when there are such signs, prompt euthanasia is recommended. It is recommended to discourage mAb production by the mouse ascites method, but it should be permitted when it is scientifically justified and approved by the relevant authorities. There has been considerable research on in vitro methods for growing hybridomas and these newer methods are less expensive, are faster, and produce antibodies in higher concentration than has been the case in the past (see below). This technique requires some expertise, requires special media, and can be expensive and time consuming. The alternative is to grow hybridoma cells in a tissue-culture medium. If the in vivo method produces pain or distress in animals, regulations call for a search for alternatives. However, if too much fluid accumulates or if the hybridoma is an aggressive cancer, the mouse will likely experience pain or distress. The mouse ascites method is inexpensive, easy to use, and quick. When injected into a mouse, the hybridoma cells multiply and produce fluid (ascites) in its abdomen, which contains a high concentration of antibody. This can be done either in vivo or in vitro: injecting hybridoma cells into the peritoneal cavity of a mouse (in vivo) or using in vitro cell-culture techniques. ![]() Once the desired hybridoma cell lines have been cloned and selected, production of the antibodies is the next stage. The created hybridoma cell lines that are producing the desired antibody are isolated and cloned to obtain the maximal yield of antibodies. The hybridoma cells will secrete the antibodies into the cell culture medium, which can be tested for the right specificity. A tumour of the fused cells is called a hybridoma, but in practice the fused cells initially grown in vitro and selected for its secreted antibodies are called hybridomas as well. Antibody-producing cells from the animal’s spleen (B-cells) are fused with a cancer cell (a myeloma) to make them grow and divide indefinitely while they continue producing antibodies (1). The traditional way of producing mAbs requires immunizing an animal, usually a mouse, a rat or a rabbit. Although, short term use polyclonal antibodies may suffice, monoclonal antibodies (mAbs) are commonly preferred when persistent use of the same antibody over time is required. IntroductionĪntibodies are important tools used by many investigators in their biomedical research and they have led to many medical advances. Large parts of the content of this paper has been ignored and outdated arguments have been taken out and replaced by contents from later publications. Washington (DC): National Academies Press (US) 1999, referred to in the text as (NCR 1999) a few times. This paper is mainly based on the publication of the National Research Council (US) Committee on Methods of Producing Monoclonal Antibodies. ![]()
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