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Year 2012 - Volume 32, Number 7
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Production and characterization of monoclonal antibodies against Campylobacter fetus subsp. venerealis, 32(7)640-644
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ABSTRACT.- Alves T.M., Heneine L.G.D., Araújo B.S., Silva L.M., Campos P.C., Hermogenes M.S. & Lage A.P. 2012 Production and characterization of monoclonal antibodies against Campylobacter fetus subsp. venerealis. Pesquisa Veterinária Brasileira 32(7)640-644. Laboratório de Bacteriologia Aplicada, Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais. Av. Antônio Carlos 6627, Cx. Postal 567, Belo Horizonte, MG 30123-970, Brazil. E-mail: alage@vet.ufmg.br.
Myeloma cells Sp2/0-Ag14 and spleen cells from BALB/c mouse immunized with sonicated Campylobacter fetus subsp. venerealis NCTC 10354 were fused with polyethylene glycol (PEG) for the selection of clones producing antibodies. Clones were obtained by limiting dilution and screened for the production of specific antibodies to C. fetus subsp. venerealis NCTC 10354 by indirect ELISA and western blot against a panel of bacteria: C. fetus subsp. venerealis NCTC 10354, C. fetus subsp fetus ADRI 1812, C. sputorum biovar sputorum LMG 6647, C. lari NCTC 11352, and Arcobacter skirrowii LMG 6621 for the ELISA and C. fetus subsp. venerealis NCTC 10354 and C. sputorum biovar sputorum LMG 6647 for the western blotting. Fifteen clones producing monoclonal antibodies (MAbs) anti-C. fetus subsp. venerealis of the IgM (1) and IgG (14) classes were further screened for species-specificity. Four clones of the 15 obtained were producers of species-specific monoclonal antibodies (MAbs): two were specific for C. fetus subsp. venerealis and two were specific for C. fetus subsp. fetus. None of the clones were reactive against C. sputorum biovar sputorum LMG 6647. All clones recognized a protein with molecular mass of approximately 148 kDa from lysed C. fetus subsp. venerealis NCTC 10354. |
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