Oligonucleotides are short single- or double-stranded nucleic acid polymers that interact with target RNA or DNA to elicit biological responses. There are currently 19 FDA approved oligonucleotide therapeutics.
Advanced analytical strategies are needed to ensure high-fidelity purification, testing and scale-up of oligonucleotides.
This infographic highlights the high-performance purification techniques that are essential for high-quality oligo production.
Download this infographic to discover:
- Analytical approaches for confident and superior oligonucleotide purification
- The role of LC and LC-MS in high-fidelity purification, testing and scale-up
- Which analytical and purification strategies are right for your oligonucleotide
Oligonucleotide therapeutic classes
18-30 bases
siRNA/ASO
Up to 200 bases
gRNA
1000s of bases
mRNA
Ion-Pair Reversed
Phase (IP-RP)
Ion-pairing reagents -
aqueous mobile phase
Non-Ion Pair Reversed
Phase (RP)
Polar interactions -
organic mobile phase
Hydrophilic Interaction
(HILIC)
Polar interactions -
aqueous mobile phase
Anion Exchange
(AEX)
Anionic interactions -
aqueous mobile phase
Ion-pairing
agent
N
δ+
Anion
(acidic)
backbone
R R R R R
O- R
B+
O O
OH
B
O O
Ẍ
OH
O O O O
Selectivity type Ion pair
reversed-phase
(IP-RP)
Non-ion pair
reversed-phase
(IP-free)
Hydrophilic
interaction
chromatography
(HILIC)
Anion
exchange
(AEX)
Oligo
classes
siRNA/ASO
gRNA
mRNA
pH and temperature
stability
Column
types
Analytical
Semi-prep
Preparative
MS compatible With desalting
Particle
type
Fully porous
Superficially
porous
Non-porous
Unlock Analytical
and Purification Strategies
for Oligonucleotides
Novel column separation chemistries to enhance performance and versatility
When analyzing oligonucleotides and other complex biological molecules, advanced
analytical strategies are needed to ensure accuracy. Liquid chromatography (LC) and liquid
chromatography mass spectrometry (LC/MS) based methods enable high-fidelity purification,
testing, and scale-up of oligonucleotides.
Agilent has innovated unique column chemistries to allow diverse analytical approaches to
enable confident and superior oligonucleotide separations.
Oligonucleotides are
short single- or doublestranded
nucleic acid
polymers that interact
with target RNA or
DNA to elicit biological
responses.
There are currently
19 FDA approved
oligonucleotide
therapeutics, including
6 small interfering
RNAs (siRNAs)
and 11 antisense
oligonucleotides
(ASOs).
High-fidelity oligo
preparations
are essential for
downstream chemical
modification to
support stability and
pharmacological
properties.
High-performance
purification techniques
are pivotal to
producing high-quality
oligos suitable for
modification and
testing, as well as scale
up to ensure high purity
yields.
Sponsored by:
Oligonucleotide chromatography separation chemistries
Gather insights on which analytical and purification
strategies are right for your oligonucleotide
Utilize guides and selection tools to help you quickly choose the right columns, standards,
reagents, and supplies for your analysis. Agilent can help you obtain the resolution and
performance you need.
Learn more:
Oligonucleotide Columns and Standards Oligonucleotide Therapeutic Solutions
View application notes
Analysis of Oligonucleotides with Ion
Exchange Chromatography and Agilent
Infinity II Bio LC System
Anion Exchange (AEX):
Evaluating HILIC Stationary Phases for
Oligonucleotide Separation by LC/MS
HILIC:
Analysis of Oligonucleotides Using an
Ion-Pairing-Free Reversed-Phase Methods
with TOF LC/MS
Non-IP-RP:
Superficially Porous Columns for Semi-
Preparative Purification of Synthetic
Oligonucleotides
Semi-Prep:
Oligonucleotide Analysis with Ion-Pair
Reversed-Phase Chromatography and
Agilent 1260 Infinity II Prime LC
IP-RP:
