Immunochemical comparison of human and rhesus monkey liver microsomal and the hepatocellular carcinoma-induced human serum epoxide hydrolases (preneoplastic antigens)

Basis for an enzyme-linked immunoabsorbent assay

David E. Moody, Bruce D. Hammock, Boris H. Ruebner, David W. Hillman, James H. Hillman

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

An antibody (anti-EH) specific for microsomal epoxide hydrolase (mEH)from rhesus monkey liver has been used to test the immunochemical relationship between human liver mEH and the serum EH levels in human patients with hepato-cellular carcinoma (HCC). Immunoblots of separated rhesus monkey and human liver microsomal proteins revealed that anti-EH was selective for a single polypeptide band of similar mol. wt, 7sim; 49 kd, in both species. Anti-EH was also able to precipitate 100% of the activity for two substrates specific in the mouse for mEH, cis-stilbene oxide and benzo[a]-pyrene-4, 5-oxide, in solubilized human liver microsomes. In contrast, only 20% of the microsomal trans-stilbene oxide hydrolase activity was precipitated under similar conditions, providing immunochemical evidence that a distinct EH, with substrate selectivity similar to the cytosolic EH, resides in human liver microsomes. Immunoprecipitation of serum from a patient with elevated EH activity resulted in total precipitation of cis-stilbene oxide hydrolase activity. An enzyme-linked immunoabsorbant assay (ELISA) was developed using anti-EH with detection limits of 1 ng/ml. A high correlation between the enzymatically and immunochemically determined levels of serum EH provided further evidence for the immunochemical similarity of human liver microsomal and serum EH. In addition, the ELISA was equally capable of identifying elevated serum EH in patients with HCC, and should prove invaluable in evaluating the effectiveness of serum EH levels as a marker for HCC.

Original languageEnglish (US)
Pages (from-to)343-349
Number of pages7
JournalCarcinogenesis
Volume10
Issue number2
DOIs
StatePublished - Feb 1989

Fingerprint

Epoxide Hydrolases
Hydrolases
Antigens
Macaca mulatta
Liver
Hepatocellular Carcinoma
Assays
Enzymes
Oxides
Serum
Liver Microsomes
Carcinoma
mouse EPHX1 protein
Substrate
Detection Limit
Polypeptides
Pyrene
Substrates
Selectivity
Immunoprecipitation

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Physiology (medical)
  • Physiology
  • Behavioral Neuroscience
  • Cancer Research

Cite this

Immunochemical comparison of human and rhesus monkey liver microsomal and the hepatocellular carcinoma-induced human serum epoxide hydrolases (preneoplastic antigens) : Basis for an enzyme-linked immunoabsorbent assay. / Moody, David E.; Hammock, Bruce D.; Ruebner, Boris H.; Hillman, David W.; Hillman, James H.

In: Carcinogenesis, Vol. 10, No. 2, 02.1989, p. 343-349.

Research output: Contribution to journalArticle

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abstract = "An antibody (anti-EH) specific for microsomal epoxide hydrolase (mEH)from rhesus monkey liver has been used to test the immunochemical relationship between human liver mEH and the serum EH levels in human patients with hepato-cellular carcinoma (HCC). Immunoblots of separated rhesus monkey and human liver microsomal proteins revealed that anti-EH was selective for a single polypeptide band of similar mol. wt, 7sim; 49 kd, in both species. Anti-EH was also able to precipitate 100{\%} of the activity for two substrates specific in the mouse for mEH, cis-stilbene oxide and benzo[a]-pyrene-4, 5-oxide, in solubilized human liver microsomes. In contrast, only 20{\%} of the microsomal trans-stilbene oxide hydrolase activity was precipitated under similar conditions, providing immunochemical evidence that a distinct EH, with substrate selectivity similar to the cytosolic EH, resides in human liver microsomes. Immunoprecipitation of serum from a patient with elevated EH activity resulted in total precipitation of cis-stilbene oxide hydrolase activity. An enzyme-linked immunoabsorbant assay (ELISA) was developed using anti-EH with detection limits of 1 ng/ml. A high correlation between the enzymatically and immunochemically determined levels of serum EH provided further evidence for the immunochemical similarity of human liver microsomal and serum EH. In addition, the ELISA was equally capable of identifying elevated serum EH in patients with HCC, and should prove invaluable in evaluating the effectiveness of serum EH levels as a marker for HCC.",
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