Expression of human aspartyl beta-hydroxylase and preparation of its monoclonal antibody / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 659-666, 2011.
Artículo
en Chino
| WPRIM
| ID: wpr-324515
ABSTRACT
We investigated the mechanism of human aspartyl beta-hydroxylase (HAAH) in early diagnosis of tumors. The encoding gene of HAAH was cloned from the hepatic carcinoma by RT-PCR and expressed as a fused protein in the prokaryotic vector pBV-IL1. The expressed HAAH was purified by Ni(2+)-NTA purification column and the purified protein was then used to immunize Balb/c mice. Three hybridoma cell lines (respectively designated H3/E10, E4/F12 and G4/D8) stably expressing the monoclonal antibody specific to HAAH fusion protein were obtained. The specificity and sensitivity of the monoclonal antibody were assessed by indirect enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Finally, the monoclonal antibody expressed by H3/E10 cell line was used to detect the expression of HAAH in several tumor cell lines by indirect immuno-fluorescence, and the specific fluorescence was observed. In conclusion, this study successfully constructed the recombinant prokaryotic vector pBV-IL1-HAAH and prepared HAAH-specific monoclonal antibody for further study of the structure and function of the protein. The result may also lay solid foundation for the research of the molecular mechanism of HAAH in early diagnosis of tumors.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Patología
/
Proteínas Recombinantes
/
Inmunización
/
Clonación Molecular
/
Línea Celular Tumoral
/
Alergia e Inmunología
/
Vectores Genéticos
/
Genética
/
Hibridomas
/
Oxigenasas de Función Mixta
Tipo de estudio:
Estudio de tamizaje
Límite:
Animales
/
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2011
Tipo del documento:
Artículo
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