Purification and isotyping of monoclonal antibodies against some important fruit diseases
1990
Espino, T.M. | Exconde, S.B. | Maldo, S.M. | Bite, M.F.P. | Zipagan, F.B. (Philippines Univ., Los Banos, College, Laguna (Philippines). National Inst. of Biotechnology and Applied Microbiology)
Purification of monoclonal antibodies against citrus trizteza virus (CTV), papaya ring spot virus (PRSV), banana bunchy top virus (BBTV) inclusions, and leaf mottling bacteria (LMB) was done by affinity chromatography on protein A-Sepharose. The unbound immunoglobulins which consisted of IgM, Iga and IgG3 were further purified by gel filtration using Biogel A-5M column. Fractions from this purification will be further characterized and identified. Results showed that IgG2a existed in all the cell lines tested. The isotype or antibody class of each monoclone was determined by the Ouchterlony immunodiffusion technique using their respective supernatant. Identification of the class of the monoclonal antibodies allows us to select and propagate clones of the desired immunoglobulin type. Results of the test showed that most of the cell lines tested were of the IgM class.
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