Versatile and Selective Biomolecule Pulldown with Combinatorial DNA-Crosslinked Polymers

Area of Science:

• Biotechnology
• Molecular Biology
• Biochemistry

Background:

• Current biomolecule isolation methods are expensive, prone to off-target binding, and lack flexibility.
• There is a need for efficient, cost-effective, and versatile platforms for capturing diverse biomolecules.

Purpose of the Study:

• To introduce LASSO (crossLink-Assisted Sequence-Selective isOlation), a novel platform for sequence-selective biomolecule isolation.
• To demonstrate LASSO's efficiency, capacity, and versatility across various biomolecules and workflows.

Main Methods:

• LASSO utilizes programmable polymer phase separation triggered by combinatorial crosslinker libraries.
• The platform forms swollen polymer agglomerates with minimal background binding for biomolecule capture.
• Switchable aptamers and gentle release mechanisms facilitate specific capture and recovery of target molecules.

Main Results:

• LASSO achieved >80% pulldown efficiency for DNA, SARS-CoV-2 RNA, and human thrombin.
• Demonstrated 8-20x higher binding capacity compared to commercial microbeads (4 nmol mg-1 polymer).
• In RNA-seq, LASSO achieved 86% ribosomal RNA depletion with significantly fewer off-target outliers than existing methods. Captured thrombin retained 98% enzymatic activity post-isolation.

Conclusions:

• LASSO provides a highly efficient, cost-effective, and stable platform for biomolecule isolation.
• Its modularity and ease of use position it to advance diagnostics, transcriptomics, and bionanotechnology.
• LASSO offers a superior alternative to current methods, reducing costs from $46-$51 to $0.96 per sample.