Automated detection of anti-double-stranded DNA antibody in systemic lupus erythematosus serum by flow immunoassay

Tae Kyu Lim, Yumiko Komoda, Noriyuki Nakamura, Tadashi Matsunaga

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We describe a novel automated flow immunoassay system for quantification of anti-double-stranded (ds) DNA autoimmune antibodies in the serum of patients suffering from systemic lupus erythematosus, dsDNA (360 bp) was covalently coupled with alkaline phosphatase (ALP) to form a novel analytical reagent (ALP-DNA). After immunoreaction, antibody-antigen complexes between ALP-DNA and anti-dsDNA monoclonal antibody were separated from unreacted ALP- DNA by an ion-exchange column on the basis of the difference in isoelectric point. Antibody-antigen complexes were subsequently quantified by luminescence following addition of 3-(2'-spiroadamantane)-4-methoxy-4-(3- phosphoryloxy)phenyl-1,2-dioxetane. The assay yielded a linear relationship between signal and concentration of anti-dsDNA monoclonal antibody in the range of 0-300 μg/mL. This simple technique permits the assay of anti-dsDNA autoimmune antibodies within 25 min. The ion-exchange column was simply regenerated by occasional elution with eluent (20 mM N-methylpiperazine, pH 5.5) supplemented with 0.5 M NaCl, to remove unreacted ALP-DNA.

Original languageEnglish
Pages (from-to)1298-1302
Number of pages5
JournalAnalytical Chemistry
Volume71
Issue number7
DOIs
Publication statusPublished - 1999 Apr 1
Externally publishedYes

Fingerprint

Alkaline Phosphatase
Antibodies
DNA
Antigen-Antibody Complex
Assays
Ion exchange
Monoclonal Antibodies
Luminescence

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Automated detection of anti-double-stranded DNA antibody in systemic lupus erythematosus serum by flow immunoassay. / Lim, Tae Kyu; Komoda, Yumiko; Nakamura, Noriyuki; Matsunaga, Tadashi.

In: Analytical Chemistry, Vol. 71, No. 7, 01.04.1999, p. 1298-1302.

Research output: Contribution to journalArticle

@article{bef526d0c1a84fe584eba2b4b22e6d78,
title = "Automated detection of anti-double-stranded DNA antibody in systemic lupus erythematosus serum by flow immunoassay",
abstract = "We describe a novel automated flow immunoassay system for quantification of anti-double-stranded (ds) DNA autoimmune antibodies in the serum of patients suffering from systemic lupus erythematosus, dsDNA (360 bp) was covalently coupled with alkaline phosphatase (ALP) to form a novel analytical reagent (ALP-DNA). After immunoreaction, antibody-antigen complexes between ALP-DNA and anti-dsDNA monoclonal antibody were separated from unreacted ALP- DNA by an ion-exchange column on the basis of the difference in isoelectric point. Antibody-antigen complexes were subsequently quantified by luminescence following addition of 3-(2'-spiroadamantane)-4-methoxy-4-(3- phosphoryloxy)phenyl-1,2-dioxetane. The assay yielded a linear relationship between signal and concentration of anti-dsDNA monoclonal antibody in the range of 0-300 μg/mL. This simple technique permits the assay of anti-dsDNA autoimmune antibodies within 25 min. The ion-exchange column was simply regenerated by occasional elution with eluent (20 mM N-methylpiperazine, pH 5.5) supplemented with 0.5 M NaCl, to remove unreacted ALP-DNA.",
author = "Lim, {Tae Kyu} and Yumiko Komoda and Noriyuki Nakamura and Tadashi Matsunaga",
year = "1999",
month = "4",
day = "1",
doi = "10.1021/ac980899r",
language = "English",
volume = "71",
pages = "1298--1302",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "7",

}

TY - JOUR

T1 - Automated detection of anti-double-stranded DNA antibody in systemic lupus erythematosus serum by flow immunoassay

AU - Lim, Tae Kyu

AU - Komoda, Yumiko

AU - Nakamura, Noriyuki

AU - Matsunaga, Tadashi

PY - 1999/4/1

Y1 - 1999/4/1

N2 - We describe a novel automated flow immunoassay system for quantification of anti-double-stranded (ds) DNA autoimmune antibodies in the serum of patients suffering from systemic lupus erythematosus, dsDNA (360 bp) was covalently coupled with alkaline phosphatase (ALP) to form a novel analytical reagent (ALP-DNA). After immunoreaction, antibody-antigen complexes between ALP-DNA and anti-dsDNA monoclonal antibody were separated from unreacted ALP- DNA by an ion-exchange column on the basis of the difference in isoelectric point. Antibody-antigen complexes were subsequently quantified by luminescence following addition of 3-(2'-spiroadamantane)-4-methoxy-4-(3- phosphoryloxy)phenyl-1,2-dioxetane. The assay yielded a linear relationship between signal and concentration of anti-dsDNA monoclonal antibody in the range of 0-300 μg/mL. This simple technique permits the assay of anti-dsDNA autoimmune antibodies within 25 min. The ion-exchange column was simply regenerated by occasional elution with eluent (20 mM N-methylpiperazine, pH 5.5) supplemented with 0.5 M NaCl, to remove unreacted ALP-DNA.

AB - We describe a novel automated flow immunoassay system for quantification of anti-double-stranded (ds) DNA autoimmune antibodies in the serum of patients suffering from systemic lupus erythematosus, dsDNA (360 bp) was covalently coupled with alkaline phosphatase (ALP) to form a novel analytical reagent (ALP-DNA). After immunoreaction, antibody-antigen complexes between ALP-DNA and anti-dsDNA monoclonal antibody were separated from unreacted ALP- DNA by an ion-exchange column on the basis of the difference in isoelectric point. Antibody-antigen complexes were subsequently quantified by luminescence following addition of 3-(2'-spiroadamantane)-4-methoxy-4-(3- phosphoryloxy)phenyl-1,2-dioxetane. The assay yielded a linear relationship between signal and concentration of anti-dsDNA monoclonal antibody in the range of 0-300 μg/mL. This simple technique permits the assay of anti-dsDNA autoimmune antibodies within 25 min. The ion-exchange column was simply regenerated by occasional elution with eluent (20 mM N-methylpiperazine, pH 5.5) supplemented with 0.5 M NaCl, to remove unreacted ALP-DNA.

UR - http://www.scopus.com/inward/record.url?scp=0033119589&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033119589&partnerID=8YFLogxK

U2 - 10.1021/ac980899r

DO - 10.1021/ac980899r

M3 - Article

VL - 71

SP - 1298

EP - 1302

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 7

ER -