Hybridoma technology

Volume 5 In Vitro Immunization in Hybridoma Technology (Borrebaeck, Editor)... 

Volume 121. Immunochemical Techniques (Part I Hybridoma Technology and... 

Generation of antibodies that can recognize and bind to specific viruses is straightforward. A sample of live or attenuated virus, or a purified component of the viral caspid, can be injected into animals to stimulate polyclonal antibody production (or to facilitate monoclonal antibody production by hybridoma technology). Harvested antibodies are then employed to develop specific immunoassays that can be used to screen test samples routinely for the presence of that specific virus. Immunoassays capable of detecting a wide range of viruses are available commercially. The sensitivity, ease, speed and relative inexpensiveness of these assays render them particularly attractive. 

Hybridoma technology (Chapter 13) facilitated development of immunoassays capable of detecting and quantifying cytokines. 

The basis of monoclonal antibody production by hybridoma technology... 

The practical achievement of this goal was held up for 18 years, primarily because of the great difficulty in isolation and purification of single-species proteins from the immune repertoire. During that time, many attempts to elicit catalysis by inhomogeneous (i.e. polyclonal) mixtures of antibodies were made and failed (e.g. Raso and Stollar, 1975 Summers, 1983). The problem was resolved in 1976 by Kbhler and Milstein s development of hybridoma technology, which has made it possible today both to screen rapidly the complete immune repertoire and to produce in vitro relatively large amounts of one specific monoclonal antibody species (Kohler and Milstein, 1975 Kohler et al., 1976). 

The next development was the production of monoclonal antibodies (MAbs) in the mid-1970s. This uses hybridoma technology, which involves the fusion of antibody-producing B cells to immortal myeloma cells. Figure 4.4 shows the preparation of MAbs using hybridoma techniques. A more detailed discussion of biopharmaceuticals production is presented in Section 10.5. 

A further advance in antibody technology is the development of transgenic mouse human strains. XenoMouse animals have been engineered in such a way that they now produce exclusively human antibodies rather than murine antibodies when immunized. The use of XenoMouse animals to produce MAbs avoids the need for any engineering of the antibody genes, since the products are already 100% human protein. XenoMouse animals are fully compatible with standard hybridoma technology and can be readily adopted by laboratories experienced in monoclonal antibody production [56]. 

The invention of B cell hybridoma technology (K3) has allowed the generation of various kinds of useful antibodies, even very minor or rare antibody species elicited in serum by the conventional procedure, as a pure immunochemical reagent in almost unlimited amounts. Among such new-generation monoclonal antibodies, anti-idiotype antibodies and anti-immune complex (anti-metatype) antibodies have been successfully introduced as key reagents enabling noncompetitive hapten immunoassays (Fig. 11). 

The development of hybridoma technology by Milstein and Kohler in 1975 revolutionized the antibody field and radically increased the purity and specificity of antibodies used in the clinic and for diagnostic tests in the laboratory. Hybridomas consist of antibody-forming cells fused to immortal plasmacytoma cells. Hybrid cells that are stable and produce the required antibody can be subcloned for mass culture for antibody production. Large-scale fermentation facilities are now used for this purpose in the pharmaceutical industry. 

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