A new model ELISA, based on two monoclonal antibodies, for quantification of fatty acid synthase.
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.
Wang
Young Y
YY
Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Kuhajda
Francis P
FP
Cheng
Paul
P
Chee
Wey Yeeng
WY
Li
Tianwei
T
Helzlsouer
Kathy J
KJ
Sokoll
Lori J
LJ
Chan
Daniel W
DW
D000328:Adult; D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor / Q000032:analysis / Q000097:blood; D001943:Breast Neoplasms / Q000201:enzymology; D016022:Case-Control Studies; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D064429:Fatty Acid Synthases / Q000032:analysis* / Q000097:blood / Q000276:immunology; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D064429:Fatty Acid Synthases