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

J Immunoassay Immunochem. 2002;23(3):279-92. doi: 10.1081/IAS-120013027.

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 3.22 ng/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.

Publication types

  • Comparative Study
  • Validation Study

MeSH terms

  • Adult
  • Antibodies, Monoclonal*
  • Biomarkers, Tumor / analysis
  • Biomarkers, Tumor / blood
  • Breast Neoplasms / enzymology
  • Case-Control Studies
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Enzyme-Linked Immunosorbent Assay / standards
  • Fatty Acid Synthases / analysis*
  • Fatty Acid Synthases / blood
  • Fatty Acid Synthases / immunology
  • Female
  • Humans
  • Male
  • Middle Aged
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Biomarkers, Tumor
  • Fatty Acid Synthases