New high mobility group box 1 assay system

Shingo Yamada, Keiko Yakabe, Junichi Ishii, Hitoshi Imaizumi, Ikuro Maruyama

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Background: High-sensitivity sandwich ELISA methods have been developed using chemiluminescent substrates. HMGB1 (high mobility group box 1) protein has been shown to play a critical role in several inflammatory diseases and it may be involved in the development of atherosclerosis. Methods: Anti-human HMGB1 monoclonal antibodies and anti-peptide polyclonal antibodies against the peptide sequence (KPDAAKKGVVKAEK) with high antigenicity and different from the sequence of HMGB2 were developed, and the antibodies were used to construct sandwich ELISA methods with a chromogenic substrate (TMBZ) and a chemiluminescent substrate (PS-atto). Highly purified human HMGB1 was used as a standard material and high-sensitivity CRP was measured to compare with HMGB1. Results: The analytical characteristics of the ELISA method we developed were validated inter-assay and intra-assay CVs were < 10%, and the detection limit was 0.3 μg/l by the chemiluminescent method and 1 μg/l with the chromogenic substrates. HMGB1 was detected in the serum of patients with acute coronary syndrome (ACS). When a cut-off of 0.6 μg/l HMGB1 upon admission to the intensive care unit (ICU) was used, the risk of developing an acute cardiac event within 1 month after discharge of ACS patients with an abnormal HMGB1 was significantly higher than for the patients with normal values (P < 0.0001). The usefulness of HMGB1 as an acute prognostic marker was suggested. Conclusions: The assay is easy to perform and suitable for use in the hospital laboratory and for screening large populations. HMGB1 is detectable in the serum of ACS patients and that the serum concentration of HMGB1 may be a prognostic indicator in ACS patients.

Original languageEnglish
Pages (from-to)173-178
Number of pages6
JournalClinica Chimica Acta
Volume372
Issue number1-2
DOIs
Publication statusPublished - 01-10-2006

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Chromogenic Compounds
Assays
Acute Coronary Syndrome
HMGB2 Protein
HMGB1 Protein
Enzyme-Linked Immunosorbent Assay
Intensive care units
Peptides
Antibodies
Substrates
Serum
Screening
Hospital Laboratories
Monoclonal Antibodies
Intensive Care Units
Limit of Detection
Atherosclerosis
Reference Values
Population
2,4,5-trimethoxybenzaldehyde

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Yamada, S., Yakabe, K., Ishii, J., Imaizumi, H., & Maruyama, I. (2006). New high mobility group box 1 assay system. Clinica Chimica Acta, 372(1-2), 173-178. https://doi.org/10.1016/j.cca.2006.04.016
Yamada, Shingo ; Yakabe, Keiko ; Ishii, Junichi ; Imaizumi, Hitoshi ; Maruyama, Ikuro. / New high mobility group box 1 assay system. In: Clinica Chimica Acta. 2006 ; Vol. 372, No. 1-2. pp. 173-178.
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Yamada, S, Yakabe, K, Ishii, J, Imaizumi, H & Maruyama, I 2006, 'New high mobility group box 1 assay system', Clinica Chimica Acta, vol. 372, no. 1-2, pp. 173-178. https://doi.org/10.1016/j.cca.2006.04.016

New high mobility group box 1 assay system. / Yamada, Shingo; Yakabe, Keiko; Ishii, Junichi; Imaizumi, Hitoshi; Maruyama, Ikuro.

In: Clinica Chimica Acta, Vol. 372, No. 1-2, 01.10.2006, p. 173-178.

Research output: Contribution to journalArticle

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T1 - New high mobility group box 1 assay system

AU - Yamada, Shingo

AU - Yakabe, Keiko

AU - Ishii, Junichi

AU - Imaizumi, Hitoshi

AU - Maruyama, Ikuro

PY - 2006/10/1

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N2 - Background: High-sensitivity sandwich ELISA methods have been developed using chemiluminescent substrates. HMGB1 (high mobility group box 1) protein has been shown to play a critical role in several inflammatory diseases and it may be involved in the development of atherosclerosis. Methods: Anti-human HMGB1 monoclonal antibodies and anti-peptide polyclonal antibodies against the peptide sequence (KPDAAKKGVVKAEK) with high antigenicity and different from the sequence of HMGB2 were developed, and the antibodies were used to construct sandwich ELISA methods with a chromogenic substrate (TMBZ) and a chemiluminescent substrate (PS-atto). Highly purified human HMGB1 was used as a standard material and high-sensitivity CRP was measured to compare with HMGB1. Results: The analytical characteristics of the ELISA method we developed were validated inter-assay and intra-assay CVs were < 10%, and the detection limit was 0.3 μg/l by the chemiluminescent method and 1 μg/l with the chromogenic substrates. HMGB1 was detected in the serum of patients with acute coronary syndrome (ACS). When a cut-off of 0.6 μg/l HMGB1 upon admission to the intensive care unit (ICU) was used, the risk of developing an acute cardiac event within 1 month after discharge of ACS patients with an abnormal HMGB1 was significantly higher than for the patients with normal values (P < 0.0001). The usefulness of HMGB1 as an acute prognostic marker was suggested. Conclusions: The assay is easy to perform and suitable for use in the hospital laboratory and for screening large populations. HMGB1 is detectable in the serum of ACS patients and that the serum concentration of HMGB1 may be a prognostic indicator in ACS patients.

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Yamada S, Yakabe K, Ishii J, Imaizumi H, Maruyama I. New high mobility group box 1 assay system. Clinica Chimica Acta. 2006 Oct 1;372(1-2):173-178. https://doi.org/10.1016/j.cca.2006.04.016