JaLCDOI 10.18926/AMO/31686
FullText URL fulltext.pdf
Author Ohya, Shogen| Mizuno, Motowo| Kawada, Mikihiro| Nasu, Junichirou| Okada, Hiroyuki| Shimomura, Hiroyuki| Yamamoto, Kazuhide| Fujita, Teizou| Tsuji, Takao|
Abstract <p>We have previously developed an enzyme-linked immunosorbent assay (ELISA) to measure stool decay-accelerating factor (DAF) and found that stool DAF concentrations were significantly elevated in patients with colorectal cancer, suggesting that the measurement of stool DAF may be valuable for the detection of colorectal cancer. In order to refine the assay for the measurement of stool DAF, we investigated 1) effects of centrifugation of stool samples, 2) effects of detergents, and 3) adequate combination of various anti-DAF monoclonal antibodies for the ELISA system using only monoclonal antibodies. We found that high-speed centrifugation could be omitted and that only the removal of large undigested food residues by centrifugation of short duration in a low-speed benchtop microcentrifuge sufficed to adequately prepare the stool samples. Addition of 2 detergents, octyl beta-glucoside and sodium deoxycholate, known to solubilize glycosyl-phosphatidylinositol-anchored proteins such as DAF, did not influence stool DAF values. By using 2 mouse anti-DAF monoclonal antibodies (clone 4F11 and 1C6), we were able to achieve a stable ELISA for the measurement of stool DAF using a uniform source of antibodies. The results should allow us to consistently apply the DAF assay for routine use in the detection of colorectal cancer.</p>
Keywords decay-accelerating factor (DAF) colorectal cancer enzyme-linked immunosorbent assay (ELISA). monoclonal sntibodies
Amo Type Article
Published Date 2002-08
Publication Title Acta Medica Okayama
Volume volume56
Issue issue4
Publisher Okayama University Medical School
Start Page 171
End Page 176
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 12199521
Web of Science KeyUT 000177382600001