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1.
J Autoimmun ; 34(1): 59-65, 2010 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19683415

RESUMEN

A diagnostic subgroup of AIH type 1 is characterized by specific serum antibodies against soluble liver protein. The respective autoantigen was named SLA/LP/tRNP((Ser)Sec), after three homologous recombinant polypeptides were isolated from expression gene libraries. We analyzed human cultured liver cells for the human homologue of recombinant SLA/LP/tRNP((Ser)Sec) by antigen purification. In addition, a monoclonal antibody was generated against recombinant SLA-p35, a truncated recombinant SLA-reactive polypeptide. With a positive patient serum, immune affinity chromatography was performed on the 52 kD-SLA main antigenic determinant pre-enriched by ion exchange chromatography. By mass spectrometry, the 52 kD-SLA/LP/tRNP ((Ser)Sec) autoantigen was unambiguously identified in the purification product. The identity of the recombinant SLA-p35 and its human homologue was further confirmed by a specific signal of the anti SLA-p35 monoclonal antibody with purified human SLA/LP/tRNP((Ser)Sec). The 48 kD-SLA species frequently comigrating in SLA-immunoblotting however was not identified by either approach. We conclude that the native counterpart of recombinant tRNP((Ser)(Sec)) indeed is detectable with a molecular weight of 52 kD in soluble liver extract of human cells as the major antigenic component of SLA/LP/tRNP((Ser)Sec).


Asunto(s)
Anticuerpos Monoclonales/metabolismo , Autoantígenos/metabolismo , Hepatitis Autoinmune/metabolismo , Hepatocitos/metabolismo , Aminoacil-ARN de Transferencia/metabolismo , Secuencia de Aminoácidos , Autoantígenos/sangre , Autoantígenos/genética , Autoantígenos/inmunología , Extractos Celulares , Línea Celular Tumoral , Cromatografía de Afinidad , Clonación Molecular , Hepatitis Autoinmune/sangre , Hepatitis Autoinmune/diagnóstico , Hepatitis Autoinmune/inmunología , Hepatocitos/inmunología , Hepatocitos/patología , Humanos , Espectrometría de Masas , Datos de Secuencia Molecular , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , Aminoacil-ARN de Transferencia/sangre , Aminoacil-ARN de Transferencia/genética , Aminoacil-ARN de Transferencia/inmunología , Proteínas Recombinantes/sangre , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo
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