Deep Profiling of Plasma Proteoforms with Engineered Nanoparticles for Top-Down Proteomics

Area of Science:

• Proteomics
• Biochemistry
• Nanotechnology

Background:

• Human plasma proteome analysis faces dynamic range challenges, limiting detection of low-abundance proteins.
• Existing methods struggle to identify proteins beyond highly abundant ones like albumin and immunoglobins.

Purpose of the Study:

• To develop and validate a nanoparticle protein corona approach for enriching low-abundance proteins and proteoforms from human plasma.
• To significantly increase the depth of proteome analysis for identifying novel biomarkers.

Main Methods:

• Utilized a nanoparticle protein corona strategy for selective and reproducible enrichment of plasma proteins.
• Employed top-down proteomics to quantify differential enrichment and detect proteoforms.
• Analyzed 2841 proteoforms from 114 proteins to assess enrichment efficiency.

Main Results:

• Achieved up to a 10⁵-fold increase in proteome depth, detecting proteoforms from ~1 μg/mL down to ~10 pg/mL.
• Identified medium and low-abundance proteoforms involved in immunity and cell signaling, expanding beyond abundant proteins.
• Demonstrated reproducible enrichment, enhancing the applicability of proteoform research.

Conclusions:

• Nanoparticle enrichment overcomes dynamic range limitations in human plasma proteomics.
• This approach enables deeper proteoform sequencing, crucial for clinical proteomics in disease and aging studies.
• Opens new avenues for biomarker discovery by uncovering previously undetectable proteoforms.