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1.
Methods Mol Biol ; 1348: 103-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26424267

RESUMO

Polyclonal antibodies are relatively easy to produce and may supplement monoclonal antibodies for some applications or even have some advantages. The choice of species for production of (peptide) antisera is based on practical considerations, including availability of immunogen (vaccine) and animals. Two major factors govern the production of antisera: the nature of adaptive immune responses, which take place over days/weeks and ethical guidelines for animal welfare. Here, simple procedures for immunization of mice, rabbits, sheep, goats, pigs, horses, and chickens are presented.


Assuntos
Soros Imunes/imunologia , Peptídeos/imunologia , Animais , Formação de Anticorpos/imunologia , Antígenos/administração & dosagem , Antígenos/imunologia , Galinhas , Cavalos , Imunização/métodos , Imunoensaio , Camundongos , Peptídeos/administração & dosagem , Coelhos , Ovinos , Suínos
2.
Biologicals ; 40(1): 36-40, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22000164

RESUMO

Currently, an assay based on fatal sensitization of mice to histamine challenge is widely used for testing absence of residual pertussis toxin in acellular pertussis containing vaccines. For replacement of this lethal end-point assay, an alternative method based on body temperature measurement in mice has been presented, and in this study the specificity and detection limit of a dermal temperature-based assay were assessed. Test preparations containing pertussis toxin were prepared in aluminum-adjuvanted pertussis toxoid vaccine and injected intraperitoneally in histamine sensitive mice. Later the mice were challenged with histamine and the pertussis toxin-induced decrease in dermal temperature recorded. By comparison of mice treated with pertussis toxoid vaccine spiked with pertussis toxin with mice treated with pertussis toxoid vaccine alone, the assay gave a response that specifically could detect presence of pertussis toxin. The acellular pertussis containing vaccine did not interfere with the pertussis toxin-induced temperature response recorded. In tests for presence of pertussis toxin in the pertussis vaccine preparation, the detection limit of the assay was estimated to approximately 5 ng pertussis toxin per human dose of pertussis toxoid. The dermal temperature-based assay was found to be a valid method to be applied in routine quality control of vaccines.


Assuntos
Bioensaio/métodos , Contaminação de Medicamentos , Agonistas dos Receptores Histamínicos/farmacologia , Histamina/farmacologia , Toxina Pertussis , Vacina contra Coqueluche , Temperatura Cutânea/efeitos dos fármacos , Animais , Humanos , Camundongos , Toxina Pertussis/análise , Toxina Pertussis/farmacologia , Vacina contra Coqueluche/análise , Vacina contra Coqueluche/farmacologia , Sensibilidade e Especificidade
3.
Clin Toxicol (Phila) ; 47(3): 219-25, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19274499

RESUMO

UNLABELLED: BACKGROUND. Ricin is a potential bioterrorism agent and no specific antidote or treatment exists for ricin poisoning. For this reason, we developed ricin-specific antibodies that were tested in a murine model of ricin poisoning for use as antidotes against symptoms of ricin poisoning. METHODS: Mice were poisoned with a lethal dose of ricin (5 microg) and their temperature and general condition were monitored for determination of a surrogate and humane end point. Mice were then treated with injections of ricin and different combinations of polyclonal anti-ricin antibodies. Antibody effect was evaluated for various doses and using various time points from ricin to antibody injection. Also, the effect of adjuvant symptomatic treatment was examined. Brain, heart, intestines, kidney, liver, lung, pancreas, spleen, and stomach tissues were sampled for histopathological analysis. RESULTS: The mouse model was reproducible and easy to use. A clear protective effect of both anti-ricin A-chain and anti-ricin B-chain antibodies-but not of irrelevant antibodies-was demonstrated with no added effect of symptomatic treatment. CONCLUSIONS: These data suggest that specific polyclonal antibodies against ricin A- and B-chain may reproducibly protect mice against ricin poisoning, even when the antibodies are administered up to 1.5 h after poisoning.


Assuntos
Anticorpos Bloqueadores/uso terapêutico , Bioterrorismo , Intoxicação por Plantas/prevenção & controle , Ricina/imunologia , Ricina/intoxicação , Animais , Anticorpos Bloqueadores/isolamento & purificação , Especificidade de Anticorpos , Temperatura Corporal/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Feminino , Esquemas de Imunização , Imunização Passiva , Imunoglobulina G/imunologia , Camundongos , Intoxicação por Plantas/patologia , Coelhos , Reprodutibilidade dos Testes
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