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New high mobility group box 1 assay system
Shingo Yamada1, Keiko Yakabe, Junichi Ishii
1Central Institute, Shino-Test Corporation, Japan. Shingo.Yamada@shino-test.co
Summary
High mobility group box 1 (HMGB1) is detectable in acute coronary syndrome (ACS) patients. Elevated HMGB1 serum levels indicate a higher risk of cardiac events, suggesting its utility as a prognostic marker.
Area of Science:
- Biochemistry
- Immunology
- Clinical Diagnostics
Background:
- High-sensitivity sandwich ELISA methods are crucial for detecting biomarkers.
- High mobility group box 1 (HMGB1) is implicated in inflammatory diseases and atherosclerosis.
- Developing sensitive assays for HMGB1 is essential for understanding its role in disease.
Purpose of the Study:
- To develop and validate high-sensitivity sandwich ELISA methods for human HMGB1 detection.
- To evaluate HMGB1 as a potential prognostic marker in acute coronary syndrome (ACS) patients.
Main Methods:
- Development of monoclonal and polyclonal antibodies against human HMGB1.
- Construction of sandwich ELISA assays using chromogenic (TMBZ) and chemiluminescent (PS-atto) substrates.
- Validation of assay characteristics including inter/intra-assay coefficients of variation (CVs) and detection limits.
Main Results:
- The developed ELISA methods demonstrated high sensitivity with detection limits of 0.3 microg/l (chemiluminescent) and 1 microg/l (chromogenic).
- HMGB1 was detected in the serum of ACS patients.
- An HMGB1 cut-off of 0.6 microg/l predicted a significantly higher risk of cardiac events within one month post-discharge (P<0.0001).
Conclusions:
- The developed HMGB1 assay is suitable for hospital laboratory use and population screening.
- Serum HMGB1 is detectable in ACS patients.
- Serum HMGB1 concentration serves as a potential prognostic indicator for ACS patients.