Technologies & Platforms

Start Your Project Now!

Please note: Our products and services are not intended to be used directly in diagnostic or therapeutic procedures.

Here's how you can reach us...

Tel:
Email:

HPLC Platforms for Nucleic Acid Drugs

Inquiry

As the development and clinical use of nucleic acid drugs (e.g., antisense oligonucleotides, siRNA, and mRNA vaccines) increase, so does the demand for efficient and accurate analytical techniques. High-Performance Liquid Chromatography (HPLC) technology from CD Formulation addresses this need by ensuring the quality and purity of nucleic acid drugs, thereby providing essential support for quality control during both development and manufacturing.

Types of HPLC Technology Platforms

CD Formulation offers a variety of HPLC technology platforms to meet diverse analytical requirements.

Reversed Phase High-Performance Liquid Chromatography (RP-HPLC)

RP-HPLC is the most common type of chromatography, suitable for the separation and analysis of various nucleic acid molecules.

Ion Exchange High-Performance Liquid Chromatography (IEX-HPLC)

IEX-HPLC is particularly suitable for the separation of positively and negatively charged nucleic acid molecules.

Affinity-HPLC

A separation technique that utilizes specific ligands, primarily for nucleic acid molecules with distinct labels.

Size Exclusion High-Performance Liquid Chromatography (SEC-HPLC)

SEC-HPLC is a technique that separates molecules based on their size, making it suitable for the analysis of larger nucleic acids.

Sequencing HPLC

Sequencing-HPLC is used for stepwise resolution of nucleic acid sequences.

Fig.1 Types of HPLC technology platforms Fig.1 Different types of liquid chromatography analysis. (CD Formulation)

Our Solutions of HPLC Technology Platforms

CD Formulation offers an HPLC platform equipped with a variety of features that deliver comprehensive solutions for the development of customer nucleic acid formulation programs.

Analysis	Types	Descriptions
Analysis of Nucleic Acid Sequences	DNA Sequencing	Reversed-phase HPLC can be utilized to analyze the length and sequence of DNA fragments. This is typically accomplished by measuring the retention time of different fragments, which facilitates the efficient separation and analysis of Sanger sequencing fragments.
	SNP Detection	HPLC facilitates genotyping by detecting single nucleotide polymorphisms (SNPs). Nucleic acid fragments with varying sequences demonstrate distinct retention behaviors within the chromatography system, attributable to minor sequence differences.
	Mutation Detection	Used to detect point mutations and small insertion/deletion mutations in genes, mutation-amplified fragments are analyzed using reversed-phase HPLC.
Sample Purity Analysis	Oligonucleotide Purity Analysis	The purity of synthesized oligonucleotides can be determined by separating and quantifying the by-products and unreacted intermediates of the synthesis process using HPLC.
Sample Purity Analysis	Nucleic Acid Formulations Purity Analysis	Purity analysis of DNA or RNA extracted from biological samples is performed to check for the presence of contaminants or degradation products.
Quantitative Testing	Absolute Quantification of Nucleic Acids	A mixture of nucleic acids, degraded by protease, is separated using HPLC and quantified with either a UV detector or a fluorescence detector.
Quantitative Testing	Real-Time Monitoring of Enzymatic Efficiency	In the study of nuclease activity, changes in the concentrations of substrate and product are monitored in real time by HPLC to evaluate the efficiency of enzyme cleavage.
Fragment Analysis	Fragment Length Analysis	After PCR amplification, DNA fragments of varying lengths are analyzed using HPLC. This application is particularly valuable in genomic research, DNA fingerprinting, and microsatellite analysis.
RNA Analysis	MicroRNA and siRNA analysis	This method is used to isolate and purify short fragments of RNA molecules, such as microRNA (miRNA) and small interfering RNA (siRNA).
RNA Analysis	mRNA Analysis	The isolation and purification of various types of mRNA can be utilized for expression analysis and transcriptomics research.
Binding Studies	Nucleic Acid-Drug Interaction Study	This study analyzes the interactions between nucleic acids and small molecule drugs. It aims to determine the binding constants by measuring the retention times of both the drugs and nucleic acids on a chromatographic column.
Modified Nucleic Acid Analysis	Methylation Analysis	HPLC can be utilized to analyze the methylation status of DNA and RNA, such as the detection of 5-methylcytosine. Nucleic acid fragments with varying methylation states exhibit distinct retention behaviors.
Modified Nucleic Acid Analysis	Other Chemical Modifications	Various modifications, including phosphorylation and oxidation, are detected and analyzed.
Analysis of Nucleic Acid Degradation Products	Plasmid DNA Degradation Products	The purity and cleanliness of plasmid DNA are critical in gene therapy and vaccine development, and HPLC can help isolate and detect degradation products of plasmid DNA.
Analysis of Nucleic Acid Degradation Products	mRNA Vaccines	For gene or mRNA vaccines, some modified and unmodified mRNA fragments can also be detected by HPLC.

Highlights of HPLC Technology Platforms

HPLC offers exceptional resolution and sensitivity for the flexible separation and detection of nucleic acid molecules with varying lengths and modifications, facilitating accurate analysis of complex samples.
Combined with detectors such as UV, fluorescence, or mass spectrometry, HPLC offers reliable quantitative analysis and facilitates the precise identification of impurities, degradation products, and modifiers in nucleic acid drugs.
HPLC can be used for the detection of various types of nucleic acid drugs, including small interfering RNAs (siRNAs), antisense oligonucleotides, mRNA and others.
Rapid method development and efficient data processing facilitate the acquisition of validated results in a short timeframe, offering timely support for R&D as well as quality control.

Our Analytical Characterization Services

CD Formulation's advanced HPLC technology ensures accurate and reliable results to support the development and quality control of nucleic acid-based therapeutics.

Identification for Nucleic Acid Drugs

HPLC analysis confirms that the nucleic acid drug in the sample aligns with the intended target, thereby verifying both the identity and purity of the drug.

Impurity Analysis for Nucleic Acid Drugs

Our platform precisely identifies impurities in nucleic acid drugs to ensure drug safety and regulatory compliance.

Safety Analysis for Nucleic Acid Drugs

Through the HPLC platform, we can comprehensively evaluate the safety of nucleic acid drugs, including the detection of potential allergenicity and toxic metabolites.

Nucleic Acid Drug Biomarker Testing

Using HPLC technology, changes in relevant biomarkers can be accurately monitored during treatment, providing reliable data to support studies on drug efficacy and mechanisms.

Publication Data

Technology: Analysis of nucleic acid molecules by HPLC technology

Journal: Scientific reports

IF: 4.38

Published: 2019

Results:

Methods have been reported for the analysis of RNAs up to 60 nucleotides (nt), but they are inadequate for the purity assessment of RNAs up to 100 nucleotides long, which are used as single-guide RNAs (sgRNAs) in CRISPR technology and promoted as drugs. In order to obtain both characterization and purity using a single high-performance liquid chromatography (HPLC) method, the current analytical strategy is high-temperature ion-pair reversed-phase (IP-RP) chromatography in the presence of organic cosolvents. The authors demonstrated the relative stability of RNA in a denaturing/basic IEX mobile phase, developed a protocol for determining the on-column stability of any RNA, and determined the applicability of the method for sgRNA, tRNA, and mRNA quality testing.

Fig.2 Analysis of nucleic acid molecules by HPLC technology Fig.2 DNAPacRP HPLC profiles for a range of RNAs. (Kanavarioti A., 2019)

By utilizing HPLC technology, CD Formulation can assist customers in analyzing and detecting nucleic acid drugs more efficiently, thereby promoting the innovation and market application of related biopharmaceutical products. Please feel free to contact us for customized testing solutions that ensure the safety and efficacy of nucleic acid drugs.

References

Kanavarioti A. HPLC methods for purity evaluation of man-made single-stranded RNAs. Sci. Rep. 2019, 9(1): 1019.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.

Related Services