HYBRID-ON™
Biodistribution Assay

A robust, ELISA-like assay for quantifying therapeutic oligonucleotides in plasma and tissue

Start Your Project
Start Your Project

HYBRID-ON™
Biodistribution Assay

A robust, ELISA-like assay for quantifying therapeutic oligonucleotides in plasma and tissue

Start Your Project

Quantifying siRNAs and ASOs with HYBRID-ON™ Technology

Therapeutic oligonucleotides are transforming the drug development landscape—but understanding where they go, how much is delivered, and how long they persist remains a critical challenge in preclinical development. ADViRNA’s HYBRID-ON™ assay is a proprietary, ELISA-style hybridization-ligation assay that provides highly sensitive, sequence-specific detection of chemically modified siRNAs and ASOs in complex biological matrices.

With a detection range from 1 to 1000 femtomoles, HYBRID-ON enables fast, reproducible quantification of oligonucleotides in tissue lysates, plasma, and buffer extracts—without the need for mass spectrometry or custom reagents.

How It Works

Our HYBRID-ON assay uses an optimized ELSA-like workflow based on:

  • Hybridization of a capture probe to the oligo analyte
  • Enzymatic ligation to lock in probe binding
  • Fluorescence-based detection using standard plate readers

The assay is compatible with double- or single-stranded oligonucleotides, fully modified siRNAs, and conjugated formats (e.g., GalNAc, cholesterol, DCA), offering a flexible and accessible platform for PK/PD studies.

ELSA-like workflow used at ADViRNA

Key Advantages

Sensitive & Quantitative

Sensitive & Quantitative

Detects down to 1 fmol in plasma, tissue lysates, or buffer
High Specificity

High Specificity

Can distinguish between full-length siRNAs and truncated analogs differing by a single nucleotide
Flexible Input Compatibility

Flexible Input Compatibility

Validated in mouse liver lysates, FBS, human plasma, RIPA buffer, and guanidinium based RNA lysis buffers
Standardized & Scalable

Standardized & Scalable

Uses off-the-shelf reagents and requires no proprietary enzyme kits or specialty antibodies
High Throughput

High Throughput

Ideal for multi-compound in vivo biodistribution studies across

Key Advantages

Sensitive & Quantitative

Sensitive & Quantitative

Detects down to 1 fmol in plasma, tissue lysates, or buffer
Flexible Input Compatibility

Flexible Input Compatibility

Validated in mouse liver lysates, FBS, human plasma, RIPA buffer, and guanidinium based RNA lysis buffers
High Specificity

High Specificity

Can distinguish between full-length siRNAs and truncated analogs differing by a single nucleotide
Standardized & Scalable

Standardized & Scalable

Uses off-the-shelf reagents and requires no proprietary enzyme kits or specialty antibodies
High Throughput

High Throughput

Ideal for multi-compound in vivo biodistribution studies

Case Study: In Vivo Liver Detection

We validated the HYBRID-ON assay in a multi-siRNA mouse study using:

  • HTT-DCA
  • Givosiran-GalNAc
  • Inclisiran-Cholesterol

All compounds were injected simultaneously across 4 dose groups (PBS, 5, 10, 20 mg/kg), and siRNA accumulation was measured in liver lysates using HYBRID-ON alongside mRNA quantification by qPCR. 

The assay reliably quantified each compound independently and demonstrated dose-dependent accumulation consistent with pharmacologic exposure levels.

Liver representing a case study performed at ADViRNA
Graph showing different buffers performed in an ADViRNA study
Graph showing human plasma performed in an ADViRNA study
Graph showing liver extract performed in an ADViRNA study

Assay Customization

Our HYBRID-ON assay is fully customizable for your oligonucleotide sequence. We can:

  • Rapidly design and validate detection probes
  • Adapt hybridization conditions based on chemistry and delivery format
  • Run the assay on plasma, serum, or target tissue lysates

Whether you’re working with a well-characterized compound or an early-stage oligo candidate, HYBRID-ON can accelerate your PK/PD profiling and biodistribution analysis.

 If you’re developing an oligonucleotide therapeutic, HYBRID-ON gives you a powerful tool for quantifying delivery, exposure, and tissue accumulation—quickly and affordably.

Integrate HYBRID-ON Into Your Next In Vivo Study

Contact us today to discuss using our HYBRID-ON assay in your next in vivo study.

Let's Talk

Integrate HYBRID-ON Into Your Next In Vivo Study

Contact us today to discuss using our HYBRID-ON assay in your next in vivo study.

Let's Talk