Receptor-targeted delivery • discovery to preclinical

Folate–Oligonucleotide Conjugation

CDMO-ready folate conjugation for folate receptor-mediated targeting. Manufacturing support for folate‑conjugated siRNA and ASO, plus PNA and PMO—with defined 3′/5′/internal placement, custom linkers, and analytics designed for scale.

Folate receptor targeting (FRα/FRβ) siRNA, ASO, PNA, PMO compatible 3′ / 5′ / internal site control Stable or cleavable linkers Client-supplied ligands supported

Overview

Folate (vitamin B9) is a small-molecule ligand recognized by folate receptors (FRα/FRβ) that are enriched on selected cell types and many tumor models. By attaching folate to an oligonucleotide, developers can explore receptor-mediated uptake and targeted exposure in discovery and preclinical studies. 1

Folate conjugation is commonly used as a screenable targeting motif to compare ligand density, attachment site, and linker stability across in vitro uptake assays and in vivo biodistribution studies. We support stable or cleavable linkers and defined 3′/5′/internal placement for multiple oligo modalities.

Bio-Synthesis supports folate conjugation for siRNA duplexes and single-stranded modalities (ASO, PNA, PMO) with position-specific attachment and purification/analytics designed to verify conjugation completeness and product identity.

Quick facts

Folate receptor binding can enable receptor-mediated uptake; linker and placement choices influence stability, handling, and bioactivity.

FRα / FRβ

Receptor targets

Small molecule

High density ligand

3′ / 5′ / Internal
Site control
Linkers
Stable or cleavable
Key Program Highlights

Core considerations for discovery-to-preclinical folate conjugate development.

Why folateTargeting
Receptor-mediated hypothesis testing

A practical targeting handle to evaluate uptake and exposure patterns in FR-expressing systems.

DesignControl
Attachment & linker options

Defined placement (3′/5′/internal) with stable or cleavable linkers aligned to modality and mechanism.

CMCScale
Preclinical manufacturability

Purification and analytics designed to resolve unconjugated/partial conjugates and support scale-up.

Biological Context: Folate Receptor Expression

Folate receptor alpha (FRα) is overexpressed in multiple solid tumor models including ovarian, lung, breast, and certain colorectal cancers, while limited expression is observed in most normal tissues. FRβ is reported in activated macrophages and selected hematologic contexts. This differential expression pattern makes folate a practical small‑molecule ligand for receptor‑mediated targeting studies in oncology and inflammation models.

How Folate Targets Cells (Folate Receptors)

Folate receptors bind folate and can mediate endocytosis. In receptor-targeting studies, folate conjugation is used to bias uptake toward FR-expressing cells and compare design variables such as linker stability, attachment site, and ligand presentation. 2

1Binding
Receptor engagement

Folate binds folate receptors (FRα/FRβ) to initiate uptake in receptor-positive systems.

2Uptake
Endocytosis

Receptor binding can trigger internalization; linker/chemistry can influence trafficking and stability.

3Activity
Productive delivery

Oligo modality and chemistry govern where activity occurs (cytosol/nucleus) after uptake.

How folate-conjugated oligonucleotides bind folate receptors and undergo receptor-mediated endocytosis

Schematic representation of folate receptor binding, endocytosis, and intracellular delivery of folate-conjugated oligonucleotides.

Design levers to evaluate
Ligand density, attachment site (3′/5′/internal), linker stability (stable vs cleavable), and oligo chemistry (backbone/sugar/base) are common variables in folate-targeting studies.
Common Applications
AssaysUptake
Targeted Uptake Screens

Compare uptake in FR‑positive vs FR‑negative models and optimize ligand density and placement.

In vivoPK/BD
Biodistribution Studies

Profile exposure patterns and tissue distribution across linker types and conjugate architectures.

OncologyModels
Tumor Model Evaluation

Assess receptor-targeted delivery hypotheses in tumor-relevant systems with defined QC attributes.

MechanismTrafficking
Receptor‑Mediated Internalization

Study internalization/trafficking behavior and stability windows using stable vs cleavable linkers.

Supported Folate Conjugate Architectures

Folate conjugates can be configured as terminal or internal attachments and paired with modality-appropriate chemistry. Programs may evaluate single folate ligands, spacer-assisted constructs (e.g., PEG/TEG), or client-supplied folate variants when required. 2

Small moleculeFolate
Folate–oligo (single ligand)

Common format for receptor-targeting evaluation and uptake studies.

Best for: screening

SpacerPEG/TEG
Spacer-enabled folate designs

Spacer architectures can tune presentation, hydrophilicity, and handling.

Best for: developability tuning

siRNADuplex
Folate–siRNA

Folate‑conjugated siRNA with defined strand/terminal placement options, scalable duplex handling, and analytics.

Best for: gene silencing

ASOSingle-strand
Folate–ASO

Folate‑conjugated ASO (gapmer or steric‑block) with controlled attachment, impurity planning, and QC support.

Best for: antisense programs

PNA/PMOSpecialty
Folate–PNA & PMO

Conjugation tailored to backbone chemistry and downstream assay needs.

Best for: specialty modalities

ClientSupplied
Client-supplied folate variants

We can incorporate client-provided ligands or custom targeting motifs per program design.

Best for: differentiated IP

Linker Chemistry & Attachment Placement

Linker choice and attachment site influence conjugation stability, purification behavior, and biological performance. Programs often compare stable linkers versus cleavable designs depending on desired release and assay interpretation. 3

StableCMC
Stable linkers

Designed to maintain conjugate integrity during handling and circulation; supports clear interpretation of distribution.

Consider: sterics & purification

CleavableRelease
Cleavable linkers

Designed to release the active oligo under defined triggers; route selection considers stability windows.

Consider: stability window

Placement (3′ vs 5′ vs internal)

Position-specific attachment can be selected to preserve functional regions, manage duplex loading (siRNA), and align with nuclease protection and analytical strategy. We support 3′, 5′, and internal placement depending on modality and design intent.

Conjugation Chemistry Approaches

Common folate activation strategies include EDC/NHS-mediated coupling of carboxyl groups, spacer-enabled PEG or TEG linkers to improve solubility, and alternative chemistries such as click-based conjugation for defined attachment control. Chemistry selection is guided by solubility, stability, and downstream analytical resolution requirements.

Common Challenges — and How We Engineer Around Them
SolubilityHandling
Folate hydrophobicity

Spacer architectures (PEG/TEG, tuned alkyl spacing) are selected to manage aggregation and preserve handling.

PurityResolution
Close-running impurities

Purification strategies are designed to resolve unconjugated oligo, partial conjugates, and positional variants where relevant.

StabilityWindow
Linker stability trade-offs

Stable vs cleavable linkers are selected against assay intent; stability profiling supports storage and study conditions.

CMCScale
Scale translation

Defined CQAs and analytics are carried from early batches through scale-up to reduce surprises later.

Folate Conjugation Workflow

1Design
Program Design

Define modality, attachment site (3′/5′/internal), linker type, and analytical strategy.

2Synthesis
Oligo Synthesis

Solid-phase synthesis with modification control and in-process verification.

3Conjugation
Folate Coupling

Activation and covalent attachment using selected chemistry platform.

4QC
Purification & Analytics

Resolve conjugated vs unconjugated species; confirm identity via LC‑MS and chromatographic methods.

5Scale
Scale‑Up & Release

Batch documentation and scalable execution aligned to preclinical development needs.

Preclinical Manufacturing & CMC-Aligned Analytics

Folate conjugates introduce additional CQAs: conjugation completeness, linker integrity, ligand stoichiometry, and conjugate-specific impurities. A development-aligned workflow defines these attributes early and carries them across scale. 3

PurificationResolution
Purification strategy

Resolve unconjugated oligo, partially conjugated species, and positional variants where relevant.

AnalyticsIdentity
Analytical confirmation

Confirm conjugation and composition via orthogonal assays (e.g., LC‑MS and chromatography profiles).

StabilityLinker
Stability profiling

Evaluate linker/conjugate stability under storage and handling conditions to support shelf-life and shipping.

Talk to a Scientist

Tell us your modality (siRNA, ASO, PNA, PMO), target biology and model system, and preferred folate architecture (attachment site, linker type, and any client-supplied ligand requirements). We’ll help translate design intent into a manufacturing-ready conjugate plan.

Position-specific attachment Stable or cleavable linkers CMC-aligned analytics
What to include in your request
  • Oligo type (siRNA / ASO / PNA / PMO)
  • Folate ligand format (standard folate or custom / client-supplied)
  • Attachment site (3′/5′/internal) and linker preference
  • Modification pattern (if defined)
  • Target quantity and timeline

FAQ

Is folate considered a targeting ligand?

Yes. Folate (vitamin B9) is a small-molecule ligand used to engage folate receptors and explore receptor-mediated uptake in receptor-positive systems.

Which modalities can be folate-conjugated?

We support folate conjugation for siRNA, ASO, PNA, and PMO programs with defined placement and modality-appropriate chemistry.

Where can folate be attached on an oligo?

Commonly at 3′ or 5′ termini; internal placement can also be supported depending on modality, functional regions, and analytical strategy.

What QC items matter most for folate conjugates?

Conjugation completeness, linker integrity, and resolution of unconjugated or partially conjugated impurities; identity confirmation via orthogonal analytics (e.g., LC‑MS and chromatography).

Why Choose Bio-Synthesis

Trusted by biotech leaders worldwide for over 45+ years of delivering high quality, fast and scalable synthetic biology solutions.