Anti-Oligonucleotide Antibodies

Custom Antibodies to Nucleic Acids

We are experienced in working with a variety of chemistries, including:

Conventional Oligonucleotides

DNA-RNA Hybrids

Modified Oligonucleotides

Single Stranded Oligonucleotide Mixtures (Cocktails)

Single Stranded RNA (ssRNA)

Double Stranded RNA (dsRNA)

Our Oligonucleotide Antibody Production Process

Each oligonucleotide antibody production project typically includes four phases: antigen preparation, antibody development, purification, and characterization.

The below graph depicts the timeline of an oligonucleotide antibody production case study; however, your timeline can vary depending on the specific needs of your project.

Phase 1: Antigen Preparation

We work closely with you to understand the biochemical properties of the target nucleic acid antigen and are experienced working with a variety of chemistries, including:

• Conventional oligonucleotides (12-15 bases)
• DNA-RNA hybrids
• Modified oligonucleotide with phosphorothiorate (PS) and phosphorodithioate (PdiS) modified backbones
• Single stranded oligonucleotide mixtures (cocktails)
• Single stranded RNA (ssRNA)
• Double stranded RNA (dsRNA)
• Nucleotides
• Nucleosides

Because nucleic acids by themselves are notoriously poor immunogens and are difficult to use as an analyte in immunoassays, we have developed and optimized a number of different conjugation methods that enhance the immunogenicity of the target nucleic acid.

Our conjugation methods lead to highly efficient presentation of the oligonucleotide to the host immune system, resulting in antibodies that specifically recognize backbone-specific nucleic acids (e.g. phosphorothioate) and/or compositions (i.e. RNA-DNA hybrids).

In addition to serving as an immunogen, the conjugated target nucleic acid is also a highly effective reagent for screening immunoassays and clonal selection.

Phase 2: Anti-Oligonucleotide Antibody Development

For anti-oligonucleotide polyclonal antibody generation, we will recommend a host, immunization protocols, and testing strategies. We also purify the antibody, enhance specificity by cross adsorption, and validate performance using predetermined criteria we establish with you at the start of the project.

For conventional monoclonal and recombinant antibody generation, we will recommend the clonal cell lines that yield antibody reagents with the desired performance characteristics such as sensitivity, specificity, yield, and affinity.

Phase 3: Antibody Production and Purification

We can produce and purify your anti-oligonucleotide antibody using a number of methods, depending on whether we are generating a polyclonal, conventional monoclonal, or recombinant antibody.

Rockland's production capabilities allow for variable manufacturing batch sizes, from small scale pilot batches to large scale manufacturing lots using roller bottle cultures or large animal antisera pools.

Purification options for polyclonals, monoclonals, and recombinants:

• Protein A purification
• Protein G purification
• Protein A/G purification
• Epitope-tag purification

Phase 4: Antibody Characterization and Validation

The specificity and sensitivity of anti-oligonucleotide antibodies produced at Rockland can be characterized using a number of different immunoassays, including IHC for biodistribution, IF for intracellular localization, and ELISA for anti-drug antibody (ADA) assays. When producing fit-for-purpose antibodies, we validate performance using the intended assay.