NULISA: a proteomic liquid biopsy platform with attomolar sensitivity and high multiplexing

Area of Science:

• Biochemistry
• Molecular Biology
• Proteomics

Background:

• The blood proteome is crucial for precision medicine but challenging due to low protein abundance and wide dynamic range.
• Existing assays struggle with sensitivity and multiplexing for comprehensive proteomic analysis.

Purpose of the Study:

• To develop a highly sensitive and multiplexed assay for broad proteomic analysis.
• To overcome the limitations of traditional assays in detecting low-abundance plasma proteins.

Main Methods:

• Developed NUcleic acid Linked Immuno-Sandwich Assay (NULISA™) with a dual capture/release mechanism and oligonucleotide-conjugated antibodies.
• Implemented signal attenuation for high-abundance targets using unconjugated antibodies.
• Utilized next-generation sequencing of barcoded reporter DNA for highly multiplexed quantification.

Main Results:

• NULISA™ achieved ~10,000-fold sensitivity improvement, reaching attomolar levels.
• Demonstrated superior sensitivity compared to proximity extension assays in detecting low-abundance biomarkers.
• A 200-plex NULISA successfully quantified 124 cytokines, chemokines, and other proteins.

Conclusions:

• NULISA™ offers a powerful, sensitive, and multiplexed solution for blood proteome analysis.
• The assay facilitates the detection of critical low-abundance biomarkers in autoimmune diseases and COVID-19.
• Fully automated NULISA enhances accessibility for research and diagnostic applications.