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Methods Of Biotechnology

Montreal antibodies

Методы биотехнологии
Contents of the chapter: Textbook sections:

Obtaining monoclonal antibodies

How are the antibodies formed? The immune response is a complex process of inter-cell interactions between different types of lymphoid cells with special hormones, which has led to the active synthesis and blood-based specific antibodies against this antigen. There are receptors similar to antibodies on the surface of the Limphocytes, whose interaction with antigen in a complex inter-cell complex provides an incentive to start biosynthesis of antibodies.
An antibodies for human needs start with animal immunization. After several injections of antigens in the presence of immune response stimulants in the blood serum, specific antibodies are accumulated. The antibodies are removed from the serum in the form of a g-globulin fraction, planting the blood serum with ammonium sulfate, alcohol, PEG and other substances. The antibodies that have been received contain a lot of fun squirrels. Highly purified antibodies are emitted with iono-mean chromatography.

Standard preparations are difficult to obtain because their composition depends on the species, their individual characteristics, the immunization cycle, and other low-controlled factors. At the same time, the specificity of modern biochemical analysis is very important, i.e. the ability to allocate the substance in complex multi-component environments such as blood serums, plant juices, enzymes. It's possible to use an immunochemical technique that uses antibodies that interact in a narrowly specific manner on the principle of anthigen antithelo. Such an analysis requires completely identical antibodies that are not routinely accepted.

The solution was proposed in 1975 by English scientists Georg Köler and Cesar Milstein. They have developed a method of obtaining cellular hybrids - hybrid. The hybrids are formed by merging lymphocytes from immunized animals with bone marrow cells cultivated in vitro.

Animal immunization, in response to the antigen in the mice, is intensifying the production antibodies of V-lymphocytes. These cells can only live in the master ' s organism, when converted to an artificial nutritious environment, they die. If you put an immune cage with a tumor, hybrid cells are formed that can live in artificial environments indefinitely. At the same time, they retain the ability to synthesize antibodies.

Hybrids synthesizing certain types of antibodies are selected on selective growth media. They are then placed in a cultural fluid where they multiply and form many kin cells (clon). Such clones can synthesize antibodies called monoclonal. ♪ ♪






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