Strategies to enhance protein translocation efficiency and smoothness through biological nanopores

Advancing protein analysis with enhanced nanopore translocation methods for single-molecule applications, single molecule proteomics

Background:

Accurate protein analysis is essential for advancing proteomics. Nanopore technology exhibited great promise for real-time, single-molecule protein analysis with the potential for high-throughput and cost-effective protein sequencing. However, current methods face significant challenges in controlling protein translocation dynamics through nanopores. Proteins often translocate too quickly or exhibit irregular movement patterns, making it difficult to obtain reliable and consistent measurements. Additionally, the use of strong unfolding agents, which are necessary for linear protein translocation, often destabilizes the measurement system, creating a fundamental trade-off between protein denaturation and system stability.

Technical Overview:

Northeastern researchers have developed an innovative technology that achieves a higher degree of control over the dynamics of protein translocation through biological nanopores. This system employs a combination of engineered nanopore proteins with optimized chemical environments that enable smooth, controlled protein movement while maintaining measurement stability. The approach utilizes novel buffer compositions and voltage protocols that work synergistically to regulate translocation speed and consistency. Key innovations include modified nanopore structures that provide enhanced protein-pore interactions and chemical additives that maintain system stability even in the presence of strong protein unfolding agents.

Benefits:

• Maintains measurement stability while using strong unfolding agents, solving a key limitation in current nanopore protein analysis
• Provides enhanced control over protein translocation dynamics for more reliable measurements
• Enables smooth and consistent protein movement through nanopores
• Improves signal quality and measurement reproducibility
• Reduces measurement artifacts and noise in protein analysis

Application:

• Protein Sequencing: Enabling more accurate and efficient single-molecule protein reading
• Protein Structure Analysis: Studying unfolding dynamics and conformational changes
• Biomarker Detection: High-sensitivity detection of specific proteins in biological samples
• Drug Development: Screening protein-drug interactions at the single-molecule level

Opportunity:

• License
• Research collaboration