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.
Non-Proprietary Assay (NPA) List
Aliri brings more than 30 years of experience in bioanalytical services, developing robust methods to support all stages of drug discovery and development. We offer an extensive list of validated, off-the-shelf assays you can leverage to support a range of research applications. In addition to the hundreds of assays listed below, Aliri also excels at developing customized methods to meet your specific challenges.
Coproporphyrin-I and Coproporphyrin-III
Aliri Bioanalysis presents a newly validated non-proprietary biomarker assay (NPA) for the quantitation of Coproporphyrin I and Coproporphyrin III in human plasma. This assay offers significant advancements in biomarker analysis, applicable across diagnostics, pharmaceutical research, and patient care.
By monitoring these biomarkers, particularly in early clinical development, companies can assess OATP1B1 inhibition, potentially avoiding the need for dedicated clinical drug-drug interaction studies, saving both time and money.
Learn more about the benefits of our NPA for Coproporphyrin I and Coproporphyrin III.
LC/MS/MS VS. LC/HRMS: Identifying and quantifying oligonucleotides
LC/MS/MS and LC/HRMS are formidable instruments employed in mass spectrometry for the identification and quantification of Oligonucleotides (OGNTs). Utilizing these approaches, drug developers can obtain detailed insights into the composition and structure of molecules for biotechnology and pharmaceuticals.
Considerations for choosing the right approach for your drug development project will depend on:
- What instrument you have
- Where you want to go
- How much information you want to collect in a single injection
Discover which technique is best for you.
Optimizing small molecule drug development strategies: A step by step approach to a rugged method
Creating a rugged method for the development of small molecule drugs is crucial in the quest for successful therapies. A systematic approach not only guarantees data consistency and reliability but also empowers you to make well-informed decisions, ultimately boosting project efficiency and success.
To help you navigate the complexities and uncertainties that come with advancing small molecules for drug development, we’ve outlined a robust strategy that includes 7 crucial steps.
Download our fact sheet to learn more.
A guide to the advantages and disadvantages of sample preparation for small molecules
Accurate and reliable results in the drug discovery process rely heavily on the pivotal role of sample preparation. By adopting optimized sample preparation techniques, you can significantly improve the quality of your data.
Sample preparation for small molecules involves three primary modes:
- Protein precipitation
- Supported liquid exchange / liquid/liquid extraction
- Solid phase extraction
Our comprehensive guide offers valuable insights associated with preparing small molecule samples for analysis, including advantages and disadvantages of each preparation mode.
Bioanalytical services for animal health
Aliri has more than 18 years of experience providing state-of-the-art animal health regulatory bioanalytical services to support research and development initiatives. With a team of skilled scientists, advanced technologies, and regulatory expertise, we are committed to delivering accurate and reliable data to expedite project timelines and enhance animal health.
Aliri by the numbers:
- 45 animal health clients
- 300+ studies supported
- 70 compounds
- 100+ custom assays
- 150 biological fluids and tissues
Explore our assay experience, platforms, regulations and 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.
RNAscope assay: Single cell transcriptomics for drug target discovery
When dealing with biomarkers in drug target discovery, profiling the tissue transcriptome in its spatial organization is key to understanding and predicting that tissue’s response to immunotherapy, because the tissue function within the body relies on the precise spatial organization of the cells. This is especially true when dealing with complex and heterogeneous tissues, such as tumors.
With tumors, the relationship between the cells and their environment is what ultimately shapes a patient’s fate. Applying the RNAscopeTM assay allows that evaluation of the presence of transcripts within a spatial context. This technique then ultimately can predict the tumor’s response to immunotherapy.
To further explain the impact of the RNAscope assay, this application note details a case study on a non-small-cell lung cancer (NSCLC) sample to predict its response to immunotherapy. This case study:
- Investigates the presence of the PD-L1 drug target transcript and the Granzyme B (GZMB) biomarker of clinical outcome transcript
- Explains quality control measures taken to validate the hybridization technique
Looks at the heterogenous distribution of transcripts in the NSCLC tissue section & examines the quantification of the PD-L` and GZMB to draw conclusions on the tissue’s reaction
Timeline planning tips for importing or exporting biological samples
Missed deadlines in drug development often stem from improper planning and coordination during the import and export of biological samples. Establishing and executing a proper plan for this part of the process can help ensure that your program stays on-track, while avoiding unnecessary stress such as costly delays, lost shipments, etc.
Download this infographic to learn more about this robust process:
- Permits and government agency requirements
- Human vs animal requirements
- Timelines and expectations
- Resources and process planning