Gold nanoparticle interference study during the isolation, quantification, purity and integrity analysis of RNA

Area of Science:

• Nanotechnology
• Biochemistry
• Toxicology

Background:

• Nanoparticles (NPs) interference with toxicological assays requires validation for nucleic acid analysis.
• Citrate-capped gold nanoparticles (AuNPs) are common and their interaction with RNA needs thorough investigation.

Purpose of the Study:

• To investigate the interference of AuNPs with routine nucleic acid isolation, quantification, integrity, and purity analyses.
• To assess the impact of AuNPs on RNA spectral properties and integrity.

Main Methods:

• BEAS-2B cells were treated with AuNPs, or RNA was spiked with AuNPs pre- or post-isolation.
• Absorbance spectroscopy (220-340 nm and 190-220 nm) and electrophoresis were used to assess RNA.
• Hyperspectral imaging was employed to detect AuNPs in isolated RNA.

Main Results:

• AuNPs caused concentration-dependent decreases in absorbance (220-340 nm) and altered traditional RNA purity ratios (260/280 nm).
• Electrophoresis could not differentiate AuNP-exposed RNA integrity.
• Spectral shifts (190-220 nm) indicated structural RNA changes, with co-isolation and cell treatment yielding similar results.

Conclusions:

• AuNPs interfere with standard RNA quantification and purity assays, necessitating method validation.
• Full spectral analysis is crucial for accurate RNA assessment in the presence of AuNPs.
• Structural RNA changes occur even without detectable AuNPs in the final product, highlighting the need for advanced analytical techniques.