RESUMO
The beta-adrenergic antagonist propranolol was activated through its side chain, coupled to bovine serum albumin, and injected into BALB/c mice. After fusion of the splenocytes from these immunized mice with the NS-1 myeloma cell line, two hybridomas, producing monoclonal anti-propranolol antibodies, were isolated. Clone P-49 was monospecific for propranolol, with a significant preference for the 1-stereoisomer, as compared to the d form. On the other hand, clone P-28 cross-reacted with alprenolol as well as some other beta-antagonists. Both classes of antibodies competed with A431 epidermoid carcinoma beta 2-adrenoceptors for the binding of [3H]propranolol. When ascites cells from clone P-28 were fixed with glutaraldehyde, the anti-propranolol monoclonal antibody became cell bound. These cell-bound P-28 antibodies bind propranolol and other beta-adrenergic ligands with a similar ranking order to the soluble monoclonal antibody. The cell-bound antibody displayed a 5-fold higher affinity towards 1-propranolol than the soluble monoclonal antibody. The practical implications of these findings are discussed.
Assuntos
Anticorpos Monoclonais , Propranolol/análise , Ligação Competitiva , Carcinoma de Células Escamosas , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Humanos , Cinética , Propranolol/imunologia , Propranolol/metabolismo , Radioimunoensaio , Receptores Adrenérgicos beta/metabolismo , Soroalbumina BovinaRESUMO
Simple, convenient radioimmunoassays for total triiodothyronine and total thyroxine in human serum are discribed. Ascending dry column chromatography is used to separate the free and antibody-bound hormone. The method allows for separation by absorption, without centrifugation. The use of test tubes containing pre-measured quantities of lyophilized radioactive reagent obviates unnecessary handling of radioactive solutions.
Assuntos
Tiroxina/sangue , Tri-Iodotironina/sangue , Animais , Cromatografia em Gel , Coelhos , Radioimunoensaio/métodosRESUMO
Aldehyde fixation of hybridoma cells during active production of monoclonal antibodies (MAbs) resulted in the formation of an insoluble phase because a large number of immunologically active antibody molecules were immobilized on the fixed hybridoma cells. Such MAb preparations specific to large protein molecules and small haptens were used for the development of a homogeneous immunoassay concept.