Investigating oligonucleotide hybridization at subnanomolar level by surface plasmon resonance biosensor method

Area of Science:

• Biotechnology
• Biosensor Technology
• Oligonucleotide Therapeutics

Background:

• Antisense therapy utilizes synthetic oligonucleotides (ONs) to modulate gene expression.
• Efficient screening of modified ONs is crucial for developing effective antisense drugs.
• Label-free detection methods offer advantages in speed and reduced reagent consumption.

Purpose of the Study:

• To optimize surface plasmon resonance (SPR) biosensor technology for multianalyte screening of synthetic oligonucleotides.
• To develop a rapid, direct, and low-consumption label-free method for analyzing ONs with modified internucleotide linkages.
• To assess the sensor's capability for detecting complementary ONs with high specificity and reproducibility.

Main Methods:

• Surface plasmon resonance (SPR) biosensor optimization.
• Immobilization chemistry utilizing streptavidin-biotin interaction for probe attachment.
• Monitoring of oligonucleotide hybridization through complex formation.
• Detection of synthetic oligonucleotides (ONs) with modified internucleotide linkages.

Main Results:

• Optimized SPR biosensor enables rapid, direct, and label-free screening.
• Demonstrated detection of complementary 23-mer ONs at concentrations as low as 0.1 nM.
• Achieved high specificity and reproducibility in ON detection.
• Streptavidin-biotin immobilization chemistry yielded high ligand density and hybridization efficiency.

Conclusions:

• The optimized SPR biosensor is a powerful tool for screening synthetic oligonucleotides for antisense therapy.
• The developed method is rapid, label-free, and requires low reagent consumption.
• The sensor's high sensitivity, specificity, and reproducibility support its application in drug discovery and development.