Mass Spectrometry-Based Proteomics of Poly(methylmethacrylate)-Embedded Bone

Area of Science:

• Biochemistry
• Proteomics
• Materials Science

Background:

• Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is vital for proteomic analysis of clinical samples.
• Hard tissues like bone are often preserved in poly(methyl methacrylate) (PMMA) for histology, but their proteomic potential was unknown.

Purpose of the Study:

• To investigate the feasibility of using PMMA-embedded bone samples for large-scale proteomic analysis via LC-MS/MS.
• To optimize workflows for enhanced proteome coverage in PMMA-embedded bone.

Main Methods:

• Utilized advanced prefractionation with high-pH reversed-phase liquid chromatography.
• Employed isobaric tandem mass tag labeling for quantitative proteomic analysis.
• Compared proteomic profiles and reproducibility with cryopreserved samples.

Main Results:

• Achieved proteome coverage exceeding 1000 protein identifications using optimized methods.
• Demonstrated high reproducibility for PMMA-embedded bone sample workflows.
• Identified that decalcification and analysis of solubilization fractions are not optimal for PMMA-embedded bone.

Conclusions:

• Established a foundational workflow for large-scale proteomics of PMMA-embedded bone.
• Showcased the biological applicability using human alveolar bone and iliac crest samples, revealing site-specific proteomic profiles.
• This research opens new avenues for understanding bone biology through advanced proteomic techniques.