A one-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human neutrophil elastase

T. Hoshino, C. Doai, S. Yoshida, K. Iwata, Y. Yoshihara, Y. Okada, H. Hayakawa, T. Hayakawa

Research output: Contribution to journalArticle

Abstract

A one-step sandwich enzyme immunoassay (EIA) system for human neutrophil elastase (hNE) was established with monoclonal antibodies to quantify the hNE levels in body fluids. hNE in samples was allowed to simultaneously react with both solid-phase and peroxidase-labeled antibodies. The EIA system required 75 min for the reactions (immunoreaction for 60 min and color reaction for 15 min), and the sensitivity for standard hNE was 0.2 ng/ml (4 pg/assay, 2 S.D.s above the zero point). Linearity was obtained from 0.2 400 ng/ml (4 pg 8 ng/assay). The EIA system recognized free and α1-proteinase inhibitor complexed forms of hNE, whereas the α2-macroglobulin-complexed form was not detected. The hNE levels in the synovial fluids from patients with rheumatoid arthritis were higher than those in the fluids of those with osteoarthritis or traumatic arthritis. Saliva from patients with periodontal diseases also contained significantly increased amounts of hNE compared with that of healthy subjects.

Original languageEnglish
Pages (from-to)175-181
Number of pages7
JournalConnective Tissue
Volume27
Issue number3-4
Publication statusPublished - 1995

Fingerprint

Leukocyte Elastase
Immunoenzyme Techniques
Monoclonal Antibodies
Macroglobulins
Synovial Fluid
Periodontal Diseases
Body Fluids
Saliva
Osteoarthritis
Peroxidase
Arthritis
Rheumatoid Arthritis
Healthy Volunteers
Peptide Hydrolases
Color
Antibodies

Keywords

  • α-proteinase inhibitor
  • monoclonal antibodies
  • neutrophil elastase
  • sandwich enzyme immunoassay

ASJC Scopus subject areas

  • Rheumatology

Cite this

Hoshino, T., Doai, C., Yoshida, S., Iwata, K., Yoshihara, Y., Okada, Y., ... Hayakawa, T. (1995). A one-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human neutrophil elastase. Connective Tissue, 27(3-4), 175-181.

A one-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human neutrophil elastase. / Hoshino, T.; Doai, C.; Yoshida, S.; Iwata, K.; Yoshihara, Y.; Okada, Y.; Hayakawa, H.; Hayakawa, T.

In: Connective Tissue, Vol. 27, No. 3-4, 1995, p. 175-181.

Research output: Contribution to journalArticle

Hoshino, T, Doai, C, Yoshida, S, Iwata, K, Yoshihara, Y, Okada, Y, Hayakawa, H & Hayakawa, T 1995, 'A one-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human neutrophil elastase', Connective Tissue, vol. 27, no. 3-4, pp. 175-181.
Hoshino T, Doai C, Yoshida S, Iwata K, Yoshihara Y, Okada Y et al. A one-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human neutrophil elastase. Connective Tissue. 1995;27(3-4):175-181.
Hoshino, T. ; Doai, C. ; Yoshida, S. ; Iwata, K. ; Yoshihara, Y. ; Okada, Y. ; Hayakawa, H. ; Hayakawa, T. / A one-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human neutrophil elastase. In: Connective Tissue. 1995 ; Vol. 27, No. 3-4. pp. 175-181.
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