A new model ELISA, based on two monoclonal antibodies, for quantification of fatty acid synthase

Young Y. Wang, Francis P. Kuhajda, Paul Cheng, Wey Yeeng Chee, Tianwei Li, Kathy J. Helzlsouer, Lori J. Sokoll, Daniel W. Chan

Research output: Contribution to journalArticlepeer-review

Abstract

A new model ELISA, based on two monoclonal antibodies, was developed for the quantification of fatty acid synthase (FAS). In this sandwich assay, a monoclonal antibody M6 was used as a capture on Nunc MaxiSorp ELISA/EIA Modules and another monoclonal antibody M3, labeled with biotin, was used as a detection antibody. More than 10 molecules of biotin were labeled on the anti-FAS monoclonal antibody using modified biotinylation conditions. The within- and between-run CVs were less than 10%, and the detection limit was 3.22ng/mL. Recoveries were 98.54-121.95%, averaging 106.05%. The average FAS concentration obtained from the total 55 healthy volunteers blood was 4.07 ± 1.81 ng/mL, 4.25 ± 2.14 ng/mL in women (n = 37) and 3.70 ± 0.74 ng/mL in men (n = 18). When compared with the previously developed polyclonal-monoclonal ELISA, a different pattern of FAS levels was observed in the supernatant of two cultured breast cancer cell lines in a time course study and there was no linear correlation between the two assays using 215 human blood samples. Thus, this new model FAS-ELISA could be used as an independent assay in measuring clinical samples. In summary, this monoclonal-monoclonal FAS-ELISA is sensitive, accurate, and precise in quantification of fatty acid synthase and has potential as a complementary tool in testing clinical samples.

Original languageEnglish (US)
Pages (from-to)279-292
Number of pages14
JournalJournal of Immunoassay and Immunochemistry
Volume23
Issue number3
DOIs
StatePublished - Sep 20 2002

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Medical Laboratory Technology

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