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

ASJC Scopus subject areas

  • Analytical Chemistry

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