RESUMEN
Antibody-drug conjugates utilize the targetting potential of antibodies to improve the potential of cytostatic or cytocidal drugs. One such murine monoclonal antibody, CTM01 (mCTM01), which recognizes an epitope on breast epithelial mucin, has potential for the treatment of breast and ovarian cancers. We examine in this paper the comparative properties of mCTM01 against a number of other anti-mucin antibodies. We then describe the humanization and high level re-expression of humanized CTM01 (hCTM01), a process designed to avoid the immune response to administered murine antibodies in human patients and to produce sufficient material for clinical studies. We show that the humanized form has properties superior to mCTM01 in terms of binding affinity to antigen presented on tumour cells.
Asunto(s)
Anticuerpos Monoclonales/química , Antígenos de Neoplasias/inmunología , Neoplasias de la Mama/inmunología , Inmunoterapia , Glicoproteínas de Membrana/inmunología , Mucinas/inmunología , Neoplasias Ováricas/inmunología , Ingeniería de Proteínas , Proteínas Recombinantes de Fusión/química , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/uso terapéutico , Afinidad de Anticuerpos , Neoplasias de la Mama/terapia , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Ratones , Repeticiones de Minisatélite , Datos de Secuencia Molecular , Mucina-1 , Proteínas de Neoplasias/inmunología , Neoplasias Ováricas/terapia , Fragmentos de Péptidos/inmunología , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/uso terapéuticoAsunto(s)
Factores Quimiotácticos/inmunología , Citocinas/inmunología , Pulmón/inmunología , Fibrosis Pulmonar/inmunología , Línea Celular Transformada , Células Cultivadas/efectos de los fármacos , Células Cultivadas/inmunología , Quimiocina CCL2 , Citocinas/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Fibroblastos/inmunología , HumanosRESUMEN
The ability to select for integration of plasmid DNA into the host chromosome allows the generation of stably transfected cell lines. With transfection of a selectable marker linked to a nonselectable target gene (or by cotransfection of the two unlinked genes), high-level expression of the desired gene is obtained by selecting for amplification of the selectable marker. This unit presents two systems for gene amplification and expression. The first describes the dihydrofolate reductase (DHFR) selection system while the second is based on selection of the glutamine synthetase (GS) gene. The DHFR system is probably more widely used, and results in very high levels of amplification and expression; however, the DHFR amplification process is lengthy and may require several months to isolate and characterize a stable, amplified line. In contrast, the GS system typically requires only a single round of selection for amplification to achieve maximal expression levels.