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Detection and quantification of soluble asialoglycoprotein receptor in human serum
Author(s) -
Yago Hirokazu,
Kohgo Yutaka,
Kato Junji,
Watanabe Naoki,
Sakamaki Sumio,
Niitsu Yoshiro
Publication year - 1995
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840210218
Subject(s) - monoclonal antibody , chemistry , asialoglycoprotein receptor , chromatography , microbiology and biotechnology , immunoassay , horseradish peroxidase , affinity chromatography , gel electrophoresis , western blot , antibody , polyacrylamide gel electrophoresis , biochemistry , enzyme , biology , in vitro , hepatocyte , gene , immunology
We describe the first evidence that soluble asialoglycoprotein receptors (AGPR) are present in human serum and that they are quantifiable by an enzyme‐linked immunosorbent assay (ELISA). An affinity chromatography gel immobilized with monoclonal antibodies (McAbs) against human liver AGPR was mixed with normal sera, and the bound fraction was analyzed both by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by Western blot analysis. Immunoreactive bands corresponding to 35 to 40 kd were obtained, which were lower than those of liver AGPR (41 kd and 46 kd). Soluble AGPR in human serum was able to bind to D ‐galactose–immobilized beads, indicating that soluble AGPR remained ligand‐binding activity. In order to quantify soluble AGPR, we established an ELISA using a monoclonal antibody (30220 McAb)‐immobilized microplate and horseradish peroxidase‐labeled F(ab′) 2 of another monoclonal antibody (30201 McAb). Reproducibility of intra‐ and interassay of the ELISA were 4% to 14% and 7% to 14%, respectively. Analytical recoveries ranged from 93% to 99%. The detection limit was estimated to be 0.1 μg/L. By nonparametolic analysis, a median and a 90% tile of serum AGPR level obtained from 283 normal volunteers were 0.4 μg/L and 2.4 μg/L, respectively. (H EPATOLOGY 1995;21:383–388.)