Improvements in the measurement of stool decay-accelerating factor in the detection of colorectal cancer.
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.
Department of Medicine and Medical Science, Okayama University Graduate School of Medicine and Dentistry, Japan.
Shogen
S
Ohya
Motowo
M
Mizuno
Mikihiro
M
Kawada
Junichirou
J
Nasu
Hiroyuki
H
Okada
Hiroyuki
H
Shimomura
Kazuhide
K
Yamamoto
Teizo
T
Fujita
Takao
T
Tsuji
D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal; D018958:CD55 Antigens / Q000032:analysis* / Q000276:immunology; D002498:Centrifugation; D015179:Colorectal Neoplasms / Q000737:chemistry* / Q000175:diagnosis*; D003840:Deoxycholic Acid / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005243:Feces / Q000737:chemistry*; D005260:Female; D005960:Glucosides / Q000494:pharmacology; D006801:Humans; D008297:Male; D008875:Middle Aged
D000911:Antibodies, Monoclonal; D018958:CD55 Antigens; D003902:Detergents; D005960:Glucosides; D003840:Deoxycholic Acid