[Preparation and characterization of polyclonal antibody against Ia-associated invariant chain of Muscovy Duck (Cairina moschata)].

Liu, Shengjie; Zhu, Maoying; Chen, Deyu; Wu, Chao; Ni, Qingsheng; Yu, Weiyi

Liu, Shengjie; Zhu, Maoying; Chen, Deyu; Wu, Chao; Ni, Qingsheng; Yu, Weiyi.

Afiliación

Liu S; School of Life Sciences, Fuyang Normal College, Fuyang 236037; Anhui Key Laboratory of Zoonoses, Anhui Agricultural University, Hefei 230036, China.
Zhu M; School of Life Sciences, Fuyang Normal College, Fuyang 236037, China.
Chen D; School of Life Sciences, Fuyang Normal College, Fuyang 236037, China.
Wu C; Anhui Key Laboratory of Zoonoses, Anhui Agricultural University, Hefei 230036, China.
Ni Q; Anhui Key Laboratory of Zoonoses, Anhui Agricultural University, Hefei 230036, China.
Yu W; Anhui Key Laboratory of Zoonoses, Anhui Agricultural University, Hefei 230036, China.

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 30(6): 623-6, 2014 Jun.

Article en Zh | MEDLINE | ID: mdl-24909285

RESUMEN

OBJECTIVE: To prepare polyclonal antibody against invariant chain of Muscovy duck (Cairina moschata) (MDIi) and identify its reaction with MDIi extracted from tissues of Muscovy duck. METHODS: MDIi was amplified by PCR and used to construct the prokaryotic expression vector of pET-32a/MDIi by linking with the plasmid of pET-32a. Then pET-32a/MDIi was transformed into E.coli Rosetta to induce the prokaryotic expression. After identified by SDS-PAGE, prokaryotic expression products were further purified from running gel of SDS-PAGE and injected into mice to prepare polyclonal antibody against MDIi. The titer and specificity of the polyclonal antibody against MDIi were analyzed by indirect ELISA and Western blotting, respectively. The intensity of reaction between the polyclonal antibody and MDIi extracted from tissues of Muscovy duck was also identified by indirect ELISA. RESULTS: The prokaryotic expression vector pET-32a/MDIi was successfully constructed. About 40 kD recombinant proteins of MDIi were confirmed to be expressed in the form of inclusion body in Rosetta. Polyclonal antibody against MDIi with a titer of 1:128 000 was obtained from the immunized mice and its high specificity was demonstrated by Western blotting. The titer of reaction between the polyclonal antibody and MDIi was 1:32 000. CONCLUSION: The polyclonal antibody against MDIi was successfully prepared with a high titer and specificity. It has a strong immune reaction with MDIi extracted from tissues of Muscovy duck.

Asunto(s)

Anseriformes/inmunología; Anticuerpos Monoclonales/inmunología; Antígenos de Diferenciación de Linfocitos B/inmunología; Antígenos de Histocompatibilidad Clase II/inmunología; Proteínas Recombinantes de Fusión/inmunología; Animales; Anseriformes/genética; Anseriformes/metabolismo; Anticuerpos Monoclonales/sangre; Especificidad de Anticuerpos; Antígenos de Diferenciación de Linfocitos B/genética; Antígenos de Diferenciación de Linfocitos B/metabolismo; Western Blotting; Ensayo de Inmunoadsorción Enzimática; Escherichia coli/genética; Vectores Genéticos/genética; Antígenos de Histocompatibilidad Clase II/genética; Antígenos de Histocompatibilidad Clase II/metabolismo; Inmunización; Proteínas Recombinantes de Fusión/genética; Proteínas Recombinantes de Fusión/metabolismo

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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes de Fusión / Antígenos de Diferenciación de Linfocitos B / Antígenos de Histocompatibilidad Clase II / Anseriformes / Anticuerpos Monoclonales Tipo de estudio: Risk_factors_studies Límite: Animals Idioma: Zh Revista: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi Asunto de la revista: ALERGIA E IMUNOLOGIA Año: 2014 Tipo del documento: Article País de afiliación: China Pais de publicación: China

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