Advances in the development of a reliable assay for the measurement of stool decay-accelerating factor in the detection of colorectal cancer

Naofumi Iwagaki, Motowo Mizuno, Jyunichirou Nasu, Masakatsu Mizuno, Hiroaki Okazaki, Shinichirou Hori, Kazuhide Yamamoto, Hiroyuki Okada, Takao Tsuji, Teizo Fujita, Yasushi Shiratori

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We have previously shown that stool concentrations of decay-accelerating factor (DAF; CD55), a membrane-bound complement-regulatory protein, are significantly elevated in patients with colorectal cancer and that the measurement of stool DAF may be a valuable test for the detection of colorectal cancer. Accordingly, we are working to develop a clinically useful immunoassay for fecal DAF. A requirement for such assay is a plentiful and reliable supply of anti-DAF antibodies. We developed a sandwich enzyme-linked immunosorbent assay (ELISA) for DAF in stool specimens, using two monoclonal anti-DAF antibodies recognizing different epitopes on the DAF molecule. When we first used a biotin-labeled antibody and enzyme-linked streptavidin method, we often observed stool interference, probably due to the presence of a substance(s) with biotin activity which non-specifically bound to the Fc portion of IgG of the first anti-DAF antibody on the ELISA wells. By the use of inorganic salts in the sample-dilution buffer and HRP-labeled anti-DAF as second antibody, we circumvented the stool interference and established that the new ELISA system could reliably measure DAF at low concentrations in stool specimens. Because the new assay system uses only monoclonal antibodies, we can now consistently supply ample amounts of antibodies for routine measurement of stool DAF.

Original languageEnglish
Pages (from-to)497-507
Number of pages11
JournalJournal of Immunoassay and Immunochemistry
Volume23
Issue number4
DOIs
Publication statusPublished - 2002

Fingerprint

CD55 Antigens
Anti-Idiotypic Antibodies
Colorectal Neoplasms
Assays
Enzyme-Linked Immunosorbent Assay
Biotin
Immunosorbents
Antibodies
Monoclonal Antibodies
Streptavidin
Enzymes
Immunoassay
Epitopes
Buffers
Complement System Proteins
Salts
Immunoglobulin G
Membranes
Dilution
Molecules

Keywords

  • CD55
  • Colorectal cancer
  • Immunoassay
  • Stool

ASJC Scopus subject areas

  • Immunology
  • Pharmacology

Cite this

Advances in the development of a reliable assay for the measurement of stool decay-accelerating factor in the detection of colorectal cancer. / Iwagaki, Naofumi; Mizuno, Motowo; Nasu, Jyunichirou; Mizuno, Masakatsu; Okazaki, Hiroaki; Hori, Shinichirou; Yamamoto, Kazuhide; Okada, Hiroyuki; Tsuji, Takao; Fujita, Teizo; Shiratori, Yasushi.

In: Journal of Immunoassay and Immunochemistry, Vol. 23, No. 4, 2002, p. 497-507.

Research output: Contribution to journalArticle

Iwagaki, N, Mizuno, M, Nasu, J, Mizuno, M, Okazaki, H, Hori, S, Yamamoto, K, Okada, H, Tsuji, T, Fujita, T & Shiratori, Y 2002, 'Advances in the development of a reliable assay for the measurement of stool decay-accelerating factor in the detection of colorectal cancer', Journal of Immunoassay and Immunochemistry, vol. 23, no. 4, pp. 497-507. https://doi.org/10.1081/IAS-120015480
Iwagaki, Naofumi ; Mizuno, Motowo ; Nasu, Jyunichirou ; Mizuno, Masakatsu ; Okazaki, Hiroaki ; Hori, Shinichirou ; Yamamoto, Kazuhide ; Okada, Hiroyuki ; Tsuji, Takao ; Fujita, Teizo ; Shiratori, Yasushi. / Advances in the development of a reliable assay for the measurement of stool decay-accelerating factor in the detection of colorectal cancer. In: Journal of Immunoassay and Immunochemistry. 2002 ; Vol. 23, No. 4. pp. 497-507.
@article{478a04f1fc89426a80917c55462cb947,
title = "Advances in the development of a reliable assay for the measurement of stool decay-accelerating factor in the detection of colorectal cancer",
abstract = "We have previously shown that stool concentrations of decay-accelerating factor (DAF; CD55), a membrane-bound complement-regulatory protein, are significantly elevated in patients with colorectal cancer and that the measurement of stool DAF may be a valuable test for the detection of colorectal cancer. Accordingly, we are working to develop a clinically useful immunoassay for fecal DAF. A requirement for such assay is a plentiful and reliable supply of anti-DAF antibodies. We developed a sandwich enzyme-linked immunosorbent assay (ELISA) for DAF in stool specimens, using two monoclonal anti-DAF antibodies recognizing different epitopes on the DAF molecule. When we first used a biotin-labeled antibody and enzyme-linked streptavidin method, we often observed stool interference, probably due to the presence of a substance(s) with biotin activity which non-specifically bound to the Fc portion of IgG of the first anti-DAF antibody on the ELISA wells. By the use of inorganic salts in the sample-dilution buffer and HRP-labeled anti-DAF as second antibody, we circumvented the stool interference and established that the new ELISA system could reliably measure DAF at low concentrations in stool specimens. Because the new assay system uses only monoclonal antibodies, we can now consistently supply ample amounts of antibodies for routine measurement of stool DAF.",
keywords = "CD55, Colorectal cancer, Immunoassay, Stool",
author = "Naofumi Iwagaki and Motowo Mizuno and Jyunichirou Nasu and Masakatsu Mizuno and Hiroaki Okazaki and Shinichirou Hori and Kazuhide Yamamoto and Hiroyuki Okada and Takao Tsuji and Teizo Fujita and Yasushi Shiratori",
year = "2002",
doi = "10.1081/IAS-120015480",
language = "English",
volume = "23",
pages = "497--507",
journal = "Journal of Immunoassay and Immunochemistry",
issn = "1532-1819",
publisher = "Taylor and Francis Ltd.",
number = "4",

}

TY - JOUR

T1 - Advances in the development of a reliable assay for the measurement of stool decay-accelerating factor in the detection of colorectal cancer

AU - Iwagaki, Naofumi

AU - Mizuno, Motowo

AU - Nasu, Jyunichirou

AU - Mizuno, Masakatsu

AU - Okazaki, Hiroaki

AU - Hori, Shinichirou

AU - Yamamoto, Kazuhide

AU - Okada, Hiroyuki

AU - Tsuji, Takao

AU - Fujita, Teizo

AU - Shiratori, Yasushi

PY - 2002

Y1 - 2002

N2 - We have previously shown that stool concentrations of decay-accelerating factor (DAF; CD55), a membrane-bound complement-regulatory protein, are significantly elevated in patients with colorectal cancer and that the measurement of stool DAF may be a valuable test for the detection of colorectal cancer. Accordingly, we are working to develop a clinically useful immunoassay for fecal DAF. A requirement for such assay is a plentiful and reliable supply of anti-DAF antibodies. We developed a sandwich enzyme-linked immunosorbent assay (ELISA) for DAF in stool specimens, using two monoclonal anti-DAF antibodies recognizing different epitopes on the DAF molecule. When we first used a biotin-labeled antibody and enzyme-linked streptavidin method, we often observed stool interference, probably due to the presence of a substance(s) with biotin activity which non-specifically bound to the Fc portion of IgG of the first anti-DAF antibody on the ELISA wells. By the use of inorganic salts in the sample-dilution buffer and HRP-labeled anti-DAF as second antibody, we circumvented the stool interference and established that the new ELISA system could reliably measure DAF at low concentrations in stool specimens. Because the new assay system uses only monoclonal antibodies, we can now consistently supply ample amounts of antibodies for routine measurement of stool DAF.

AB - We have previously shown that stool concentrations of decay-accelerating factor (DAF; CD55), a membrane-bound complement-regulatory protein, are significantly elevated in patients with colorectal cancer and that the measurement of stool DAF may be a valuable test for the detection of colorectal cancer. Accordingly, we are working to develop a clinically useful immunoassay for fecal DAF. A requirement for such assay is a plentiful and reliable supply of anti-DAF antibodies. We developed a sandwich enzyme-linked immunosorbent assay (ELISA) for DAF in stool specimens, using two monoclonal anti-DAF antibodies recognizing different epitopes on the DAF molecule. When we first used a biotin-labeled antibody and enzyme-linked streptavidin method, we often observed stool interference, probably due to the presence of a substance(s) with biotin activity which non-specifically bound to the Fc portion of IgG of the first anti-DAF antibody on the ELISA wells. By the use of inorganic salts in the sample-dilution buffer and HRP-labeled anti-DAF as second antibody, we circumvented the stool interference and established that the new ELISA system could reliably measure DAF at low concentrations in stool specimens. Because the new assay system uses only monoclonal antibodies, we can now consistently supply ample amounts of antibodies for routine measurement of stool DAF.

KW - CD55

KW - Colorectal cancer

KW - Immunoassay

KW - Stool

UR - http://www.scopus.com/inward/record.url?scp=0036430004&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036430004&partnerID=8YFLogxK

U2 - 10.1081/IAS-120015480

DO - 10.1081/IAS-120015480

M3 - Article

C2 - 12458732

AN - SCOPUS:0036430004

VL - 23

SP - 497

EP - 507

JO - Journal of Immunoassay and Immunochemistry

JF - Journal of Immunoassay and Immunochemistry

SN - 1532-1819

IS - 4

ER -