Sample Preparation Strategies for Antibody-Free Quantitative Analysis of High Mobility Group Box 1 Protein

The quest to accurately quantify High Mobility Group Box 1 (HMGB1) and its redox variants in human plasma has been a challenging endeavor in biomedical research, due to the complexities presented by interference from other proteins and autoantibodies.

HMGB1, a critical inflammatory mediator and alarmin, plays a crucial role in inflammation and sickness behavior, necessitating precise quantification methods for its study.
A pioneering study has recently involved HMGBiotech’s recombinant HMGB1 for biochemistry and dsHMGB1, introducing an innovative, antibody-free sample preparation method. This approach utilizes liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) to measure HMGB1 levels without the pitfalls associated with antibody reliance, which often lead to errors in traditional assays.

The study’s findings underscore the efficacy of reversed-phase liquid chromatography for separating intact proteins in diluted plasma, achieving superior HMGB1 purification compared to immunoaffinity extraction methods. This not only signifies a methodological leap in HMGB1 analysis but also serves as a practical demonstration of the HMGB1 15N Labelled Protein’s utility in enhancing research accuracy and reliability.
By employing isotopically labeled intact HMGB1, the research showcases a novel method for its quantification, paving the way for more insightful investigations into its role across various clinical conditions. This development illustrates the critical importance of selecting high-quality research tools in achieving breakthrough findings, especially in studies involving complex biomarkers like HMGB1.

HMGBiotech Srl can provide comprehensive information to facilitate informed decision-making for research involving HMGB1.
Discover more about HMGB1 labelled for mass spectometry

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