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Soluble liver antigen: Isolation of a 35‐kd recombinant protein (SLA‐p35) specifically recognizing sera from patients with autoimmune hepatitis
Author(s) -
Volkmann Martin,
Martin Lore,
Bäurle Andrea,
Heid Hans,
Strassburg Christian P.,
Trautwein Christian,
Fiehn Walter,
Manns Michael P.
Publication year - 2001
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1053/jhep.2001.22218
Subject(s) - immunoscreening , antigen , microbiology and biotechnology , polyclonal antibodies , recombinant dna , biology , autoimmune hepatitis , virology , hepatitis , peptide sequence , gene , immunology , biochemistry , cdna library
Autoantibodies to soluble liver antigen (SLA) are considered a specific marker of autoimmune hepatitis. We have performed immunoscreening of a human liver gene expression library with an anti‐SLA—positive serum. A reactive clone with a 35‐kd open reading frame (ORF) and a 563 base pair (bp) 3′ untranslated region (UTR) was isolated (soluble liver antigen [SLA]‐p35), showing strong homology to an independently isolated putative SLA/liver‐pancreas antigen (LP) sequence (Acc. No. AF146396), and a UGA serine tRNA‐protein complex (tRNP) (Ser) Sec related protein (AJ238617), as well as different expression sequence tag (EST)‐clones from lymphatic and oncofetal tissues. Expressed in Escherichia coli , SLA‐p35 showed dose‐dependent and complete blocking of reactivity to native SLA antigen after preabsorption with the 35‐kd recombinant protein. It recognized 67/85 (78.8%) precharacterized anti‐SLA—positive sera in dilutions up to 1:40,000 in immunoblot, without detectable cross reactivity in the controls. The commercially available SLA/LP enzymelinked immunosorbent assay (ELISA), by comparison, recognized 63/85 samples (74.1%). Of the negative samples, 18% showed strong inhibition rates (80% and above) in the polyclonal inhibition ELISA. We conlude that the complementary DNA now isolated by 3 independent approaches encodes for the major but not sole antigenic component of soluble liver antigen. Although its truncated form presented here may serve to improve diagnostics based on the new recombinant polypeptide, it currently cannot fully replace the polyclonal inhibition ELISA.