RESUMEN
A murine/human chimeric antibody with specificity for Hepatitis B surface antigen has been produced by genetic engineering. The light and heavy chain variable region exons encoding the murine monoclonal antibody 2H1 were isolated and inserted into mammalian expression vectors containing the human kappa and gamma 1 constant region exons. The chimeric genes were transfected into murine Sp2/0 hybridoma cells by electroporation and transfectomas secreting chimeric antibody were isolated. Secretion levels ranged from 1-7 pg/cell/24 hr. The chimeric antibody bound specifically to Hepatitis B surface antigen and competed effectively with the parental murine monoclonal antibody for binding to these sites. Chimeric 2H1 is the first clinically relevant, genetically engineered anti-viral antibody and may represent an improved agent for the prevention of hepatitis B virus transmission.
Asunto(s)
Ingeniería Genética/métodos , Anticuerpos contra la Hepatitis B/biosíntesis , Antígenos de Superficie de la Hepatitis B/inmunología , Animales , Especificidad de Anticuerpos/genética , Secuencia de Bases , Southern Blotting , Genes de Inmunoglobulinas/genética , Humanos , Región Variable de Inmunoglobulina/genética , Ratones , Datos de Secuencia Molecular , Mapeo Restrictivo , Transfección/inmunologíaRESUMEN
Murine/human chimeric antibodies with specificity for the human TCR-alpha/beta have been produced by genetic engineering. The L and H chain V region exons encoding the murine mAb BMA 031 were isolated and inserted into mammalian expression vectors containing the human kappa and gamma 1 or gamma 4 C region exons. The chimeric genes were transfected into murine Sp2/O hybridoma cells by electroporation and transfectomas secreting chimeric antibody were isolated. Secretion levels ranged from 1 to 7 pg/cell/24 h. The chimeric antibodies bound specifically to T cells and competed effectively with the parental murine mAb for binding to these sites. The ability to promote antibody-dependent cell-mediated cytolysis was significantly enhanced in the chimeric antibodies as compared with murine BMA 031. C-dependent cytolysis, however, was not detectable with any of the antibodies. Chimeric BMA 031 is a clinically relevant, genetically engineered antibody with potential uses in transplantation, graft-vs-host disease, autoimmune diseases and other T cell-related disorders.