Phenyl SPE: A Promising Alternative to More Common Oligonucleotide Extraction
Chemical modifications made to oligonucleotides can assist in absorption, distribution, and metabolism. While many oligonucleotide-based drugs share similarities in extraction and ionization techniques, the properties of the matrix itself are often overlooked. The extraction of a variety of oligonucleotides using a standard Clarity OTX solid phase extraction plate has been more commonplace in bioanalytical laboratories. These plates have been shown to isolate RNA and DNA-based therapeutics from biological fluids and tissues in a short period of time with high recovery percentages and standard extraction buffers. Here we show an alternate extraction method for a novel antisense oligonucleotide (ASO) in spinal cord and brain tissues.
Originally, acceptable and reproducible results were obtained by using a Clarity OTX SPE plate with non-human primate cerebral spinal fluid and brain tissue homogenate. The extraction was modified in mouse brain tissue to a liquid-liquid method involving phenol-chloroform. Further optimization in spinal cord tissue, however, required a more robust method using a phenyl solid-phase extraction (SPE) plate, triethylammonium acetate buffer (TEAA), and a protein precipitation filter plate.
Antisense Oligonucleotide Extraction Optimization Reveals Significant Differences Between Non-Human Primate Spinal Cord, Mouse Spinal Cord, and Mouse Brain Tissue Homogenates
Discover the critical differences in antisense oligonucleotide (ASO) extraction methods across non-human primate spinal cord, mouse spinal cord, and mouse brain tissues. Our research highlights how unique tissue properties demand distinct extraction techniques for optimal results.
Download our poster to learn more.
Oligonucleotides: Where we started, where we stand, and where we would like to go
Oligonucleotides, which are short chains of nucleotides that make up DNA and RNA, are used in a wide range of applications in bioanalysis and play a critical role in understanding the molecular mechanisms of living cells and diseases.
Organic Synthetic Chemist, Troy Voelker PhD, discusses common methods to support the bioanalysis of oligonucleotides (OGNTs) and a future perspective regarding areas for improvement.
His insights explores a range of analytical techniques for OGNTs, including:
-Basics of oligonucleotide bioanalysis
-Selecting an approach
-Methods of extraction
-Handling Hybridization methods
-Goals for future analysis
Discover how future predictions for oligonucleotide bioanalysis will help improve medicine, therapeutics and genetic testing.