Development of the hybridoma technology has reduced the number of animals (mice, rabbits, and so on) required to produce a given antibody but with a decrease in animal welfare when the ascites method is used. Before the advent of the hybridoma method, investigators could produce only polyclonal serum antibodies this required large numbers of immunized animals and did not immortalize the antibody-producing cells, so it required repeated animal use to obtain more antibodies. It has also been possible to genetically replace much of the mouse mAb producing genes with human sequences, reducing the immunogenicity of mAb destined for clinical use in humans. Recent in vitro techniques allow the intracellular production of antigen-binding antibody fragments, but such techniques are still experimental and have an uncertain yield, efficacy, and antibody function (Frenken and others 1998). No method of generating a hybridoma that avoids the use of animals has been found. A tumor from this "immortal" cell line is called a hybridoma. The procedure yields a cell line capable of producing one type of antibody protein for a long period. The generation of mAb-producing cells requires the use of animals, usually mice. The steps in producing those cells are outlined below ( figure 1). Before production of antibodies by either method, hybrid cells that will produce the antibodies are generated. Production of monoclonal antibodies involves in vivo or in vitro procedures or combinations thereof. In: Matthews REF (ed) Diagnosis of plant virus diseases.Generation of Hybridomas: Permanent Cell Lines Secreting Monoclonal Antibodies Van Regenmortel MHV, Dubs MC (1993) Serological procedures. Association of Applied Biologists, Wellesbourne In: Jones RAC, Torrance L (eds) Developments and applications in virus testing. Van Regenmortel MHV (1986) The potential for using monoclonal antibodies in the detection of plant viruses. Torrance L, Pead MT, Larkins AP, Butcher GW (1986) Characterization of monoclonal antibodies to a UK isolate of barley yellow dwarf virus. Torrance L (1995) Use of monoclonal antibodies in plant pathology. Smith HG, Stevens M, Hallsworth PB (1991) The use of monoclonal antibodies to detect beet mild yellowing virus and beet western yellows virus in aphids. Singh RP, Sreenivasa BP, Bandyopadhyay P, Dhar PK (2004) Production and characterization of monoclonal antibodies to Peste des Petits Ruminants (PPR) virus.
Saliki JT, Libeau G, House JA, Mebus CA, Dubovi EJ (1993) Monoclonal antibody-based blocking enzyme-linked immunosorbent assay for specific detection and titration of peste-des-petits-ruminants virus antibody in caprine and ovine sera. Naidu RA, Mayo MA, Reddy SV, Jolly CA, Torrance L (1997) Diversity among the coat proteins of luteoviruses associated with chickpea stunt disease in India. Macintosh S, Robinson DJ, Harrison BD (1992) Detection of three whitefly-transmitted geminiviruses occurring in Europe by tests with heterologous monoclonal antibodies. Konaté G, Barro N, Fargette D, Swanson MM, Harrison BD (1995) Occurrence of whitefly transmitted geminiviruses in crops in Burkina Faso, and their serological detection and differentiation.
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Jordan R, Hammond J (1991) Comparison and differentiation of potyvirus isolates and identification of strain-, virus-, subgroup-specific and potyvirus group-common epitopes using monoclonal antibodies. In: Ausubel FM (ed) Current protocols in molecular biology. Ann Appl Biol 128:255–268įuller SA, Takahashi M, Hurrel JGR (2001) Fusion of myeloma cells with immune spleen cells. Phytopathology 79:869–873įranz A, Makkouk KM, Katul L, Vetten HJ (1996) Monoclonal antibodies for the detection and differentiation of Faba bean necrotic yellows virus isolates. D’Arcy CJ, Torrance L, Martin RR (1989) Discrimination among luteoviruses and their strains by monoclonal antibodies and identification of common epitopes.
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