Quantification of Serum High Mobility Group Box 1 by Liquid Chromatography/High-Resolution Mass Spectrometry: Implications for Its Role in Immunity, Inflammation, and Cancer

Overview

Journal Anal Chem

Specialty Chemistry

Date 2018 May 24

PMID 29791130

Citations 13

Authors

Liwei Weng

Lili Guo

Anil Vachani

Clementina Mesaros

Ian A Blair

Affiliations

Soon will be listed here.

Abstract

High mobility group box 1 (HMGB1) is a non-histone chromosomal protein, which can be secreted through a variety of pathways and bind to pattern recognition receptors to release pro-inflammatory cytokines. Previous studies have suggested that HMGB1 is upregulated in numerous inflammatory diseases and that it could be a biomarker for such diseases. However, these studies used immunoassay-based methods to analyze serum HMGB1. Autoantibodies to HMGB1 in serum are found in healthy control subjects as well as in patients with different diseases. HMGB1 also binds to haptoglobin, a highly abundant plasma protein. This means that antibodies used in immunoassays must compete with binding of HMGB1 to endogenous serum HMGB1 autoantibodies and haptoglobin. To overcome these potential problems, we developed and validated a specific and sensitive assay based on stable isotope dilution and immunopurification to quantify HMGB1 in plasma and serum using two-dimensional nano-ultra-high-performance liquid chromatography parallel reaction monitoring/high-resolution mass spectrometry. Using this assay, we found that serum HMGB1 in 24 healthy control subjects (6.0 ± 2.1 ng/mL) was above the mean concentration reported for 18 different diseases (5.4 ± 2.8 ng/mL) where the analyses were conducted with immunoassay methodology. In light of our finding, the role of HMGB1 in these diseases will have to be re-evaluated. The concentration of HMGB1 in citrated and EDTA-treated plasma from the same healthy control subjects was below the limit of detection of our assay (1 ng/mL), confirming that HMGB1 in serum arises when blood is allowed to clot. This means that future studies on the role of HMGB1 in vivo should be conducted on plasma rather than serum.

Citing Articles

HMGB2-induced calreticulin translocation required for immunogenic cell death and ferroptosis of cancer cells are controlled by the nuclear exporter XPO1.

Fan J, Gillespie K, Mesaros C, Blair I Commun Biol. 2024; 7(1):1234.

PMID: 39354146 PMC: 11445383. DOI: 10.1038/s42003-024-06930-y.

Ferroptosis and HMGB2 induced calreticulin translocation required for immunogenic cell death are controlled by the nuclear exporter XPO1.

Blair I, Fan J, Gillespie K, Mesaros C Res Sq. 2024; .

PMID: 38496553 PMC: 10942558. DOI: 10.21203/rs.3.rs-4009459/v2.

Quantification of human mature frataxin protein expression in nonhuman primate hearts after gene therapy.

Rojsajjakul T, Hordeaux J, Choudhury G, Hinderer C, Mesaros C, Wilson J Commun Biol. 2023; 6(1):1093.

PMID: 37891254 PMC: 10611776. DOI: 10.1038/s42003-023-05472-z.

Targeting HMGB1: A Potential Therapeutic Strategy for Chronic Kidney Disease.

Liu T, Li Q, Jin Q, Yang L, Mao H, Qu P Int J Biol Sci. 2023; 19(15):5020-5035.

PMID: 37781525 PMC: 10539693. DOI: 10.7150/ijbs.87964.

Cisplatin Dependent Secretion of Immunomodulatory High Mobility Group Box 1 (HMGB1) Protein from Lung Cancer Cells.

Gillespie K, Pirnie R, Mesaros C, Blair I Biomolecules. 2023; 13(9).

PMID: 37759736 PMC: 10526420. DOI: 10.3390/biom13091335.

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