Production and characterization of a monoclonal antibody against an Ascaris suum allergenic component
AUTOR(ES)
Pires, R.R., Oshiro, T.M., Itami, D.M., Fernandes, I., Macedo-Soares, M.F.
FONTE
Brazilian Journal of Medical and Biological Research
DATA DE PUBLICAÇÃO
2001-08
RESUMO
Ascaris suum allergenic components (PIII) separated by gel filtration chromatography of an adult worm extract were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells using polyethylene glycol (MW 1450) as fusogen. The hybridomas were cultured in HAT-containing medium and cloned at limiting dilutions. Supernatants from the growing hybrids were screened by ELISA using plates coated with PIII or the A. suum crude extract. The monoclonal antibody obtained, named MAC-3 (mouse anti-A. suum allergenic component), is an IgG1 kappa mouse immunoglobulin that specifically recognizes a 29,000 molecular weight protein (called allergenic protein) with an affinity constant of 1.7 x 10(9) M-1. The A. suum components recognized by MAC-3 induce specific IgE antibody production in immunized BALB/c mice. Ascitic fluid induced in Swiss mice by injecting ip the hybridoma cells and incomplete Freund's adjuvant was purified by affinity chromatography using a protein A-Sepharose column. The purified monoclonal antibody was then coupled to activated Sepharose beads in order to isolate the A. suum allergenic component from the whole extract by affinity chromatography.
Documentos Relacionados
- Comparison of different monoclonal antibodies against immunosuppressive proteins of Ascaris suum
- Production, Characterization, and Epitope Mapping of a Monoclonal Antibody against Aspartic Proteinase of Candida albicans
- Production and characterization of monoclonal antibodies against the lethal factor component of Bacillus anthracis lethal toxin.
- Production and characterization of monoclonal antibody against 10S DNA polymerase alpha from calf thymus.
- Characterization of an antibody directed against a surface component of normal and pleomorphic cells of Streptococcus sanguis.