Production and characterisation of monoclonal antibodies against 19-Nortestosterone / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 172-179, 2011.
Artigo
em Inglês
| WPRIM
| ID: wpr-306875
ABSTRACT
<p><b>OBJECTIVE</b>To produce anti-19-Nortestosterone (NT) monoclonal antibodies and identify their immunological characteristics.</p><p><b>METHODS</b>Hybridomas were prepared by fusing NS0 mouse myeloma cells with splenocytes isolated from immunized BALB/c mice. Noncompetitive and competitive indirect ELISA were employed to screen positive cell clones. A caprylic acid ammonium sulphate (CAAP) method was used to purify NT mAb, and the Batty saturation method was used to determine the affinity constant (Kaff).</p><p><b>RESULTS</b>Five hybridoma cell lines, named NT-1, NT-2, NT-3, NT-4, and NT-5, were identified and their corresponding mAbs were of the IgG(1) isotype with a k light chain. The Kaffs of all mAbs were between 2.6 and 4.7 × 10(9) L/mol. The titers and IC(50) values of purified ascite fluids were in the range of (0.64-2.56) × 10(5) and (0.55-1.0) ng/mL, respectively. Of all the cross-reacting steroids, (-NT was the most reactive with the mAbs at 62% with NT-1 mAb and 64% with NT-2 mAb. Negligible cross-reactivity (<0.01%) with other steroids was observed.</p><p><b>CONCLUSION</b>The establishment of these hybridomas allows the potential development of a rapid test kit, and may provide an alternative method for the detection of NT residues in food producing animals.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmocitoma
/
Kit de Reagentes para Diagnóstico
/
Baço
/
Estrutura Molecular
/
Linhagem Celular
/
Química
/
Biologia Celular
/
Alergia e Imunologia
/
Hibridomas
/
Camundongos Endogâmicos BALB C
Tipo de estudo:
Estudo prognóstico
Limite:
Animais
Idioma:
Inglês
Revista:
Biomedical and Environmental Sciences
Ano de publicação:
2011
Tipo de documento:
Artigo
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