Custom VHH Nanobody Discovery Services

 

Discover flexible nanobody discovery workflows tailored to your needs, from animal-free synthetic libraries to llama-based immune libraries, and from binder selection to engineering and expression.

 

1. Choose Your Starting Point

a. Synthetic Library Discovery

Ideal for rapid screening, high-throughput projects, modular applications, or when animal immunization is not preferred.

  • Animal-free
  • Immediate start
  • Broad binder diversity
  • Compatible with multiple target classes (proteins, peptides, toxins, etc.)

b. Immune Library Discovery

For projects that require high-affinity binders, deep epitope coverage, or involve difficult targets, we offer the development of immune VHH Nanobody libraries based on llama or alpaca immunization with your antigen.

The immunization itself is carried out by one of our trusted specialized partners, who operate professional llama and alpaca facilities. Cortalix coordinates the process closely to ensure scientific alignment, quality, and timelines.

  • Target-specific immune response
  • High natural affinity
  • Deep epitope coverage
  • Immunization coordinated by Cortalix, performed at a certified facility
  • Library construction, phage display and clone selection in-house at Cortalix

2. Selection & Enrichment (both routes)

Once your library (synthetic or immune) is in place, we proceed with tailored phage display panning to enrich for target-specific VHHs.

We optimize each round for stringency, specificity, and diversity, based on your application (diagnostic, therapeutic, etc.).

3. Clone Identification & Validation

From enriched pools, we isolate individual clones and characterize them via:

  • ELISA

Optionally, we can provide high-throughput screening of 96+ clones.

4. Downstream Engineering & Optimization

We offer additional services to tailor VHH Nanobodies to your needs:

  • CDR grafting

  • Humanization

  • Affinity maturation

  • Fc-fusion or multimerization

  • Biotinylation or other conjugations

5. Expression & Scale-up

Expression of selected VHH Nanobodies in E. coli, Pichia pastoris, or mammalian systems.

  • Small-scale expression & purification
  • Scalable production-ready formats
  • Endotoxin-free options available for in vivo use

6. Optional Add-ons

  • Diagnostic assay integration (e.g., ELISA or lateral flow)

  • Functional groups and linker conjugation

  • Fluorophore or Chelator labeling for imaging or therapy

  • Coupling to beads for analytical purposes

  • Custom formatting for analytical platforms (e.g., biosensors)

Flexibilty by Design

Our hybrid discovery model – offering both synthetic and immune VHH libraries – allows us to adapt to your project’s unique needs. Whether you require speed and modularity, or maximum affinity and epitope coverage, we help you choose the best route without compromise.

Every step is fully modular, so you can engage us from early discovery to downstream formatting, or anywhere in between.

Let’s Discuss Your Project

Not sure whether to choose synthetic or immune? We’re happy to advise based on your target and goals.

Contact us for a free consultation.

Frequently Asked Questions

Do you work with both synthetic and immune libraries?

Yes. We offer both synthetic libraries (ready-to-screen) and immune libraries based on llama or alpaca immunization with your antigen.

Can I supply my own immunized material?

Yes, if you already have PBMCs or RNA from immunized animals, we can take it from there and build the immune library.

How unique are the nanobodies that Cortalix delivers?

The libraries that Cortalix uses are highly diverse, and the likelihood of two customers receiving the same nanobody sequence is extremely small. We make every effort to ensure that each clone delivered is unique. In cases where different customers request nanobodies against the same target, we take strict measures to avoid assigning the same sequence more than once.

Do we have full ownership of the delivered nanobody sequences ?

Yes, in almost all cases you get full ownership of the nanobody DNA and amino acid sequence. If you want to use one of our specific libraries that has specific properties, such as a patented amino acid sequence in, for example, the spacer or linker, we can provide you with a non-exclusive license for worldwide commercial use. This also gives you additional advantages that your nanobody may already be protected for use by others.

Can we request only selection services?

Yes, that’s actually a very logical step to start with.
However, for any of the later steps it is often necessary to also include earlier steps, such as cloning, expressing and functionalizing nanobodies by applying specific linkers and functional groups. But just ask what the options are and we will figure it out together.

What is the typical timeline?

Synthetic library projects can start immediately and deliver binders in 4–5 weeks. Immune library projects depend on immunization timelines (typically 10–14 weeks) followed by discovery.

Do we only get the amino acid sequence at the end of STEP 1 or also the protein?

No, if desired, we also provide the DNA sequence of the nanobody itself. The amino acid (and DNA) sequence of any spacer and/or linker is not provided because this is part of an additional service (Step 5 and/or 6). If it concerns a proprietary spacer and/or linker, a non-exclusive license can be granted regarding its worldwide use.

Do you guarantee a number of unique nanobody binders from your library?

Yes, if we accept the target you supply (the protein is pure enough, large enough and can be immobilized on a 96-well plate), we guarantee the selection and delivery of at least 2 unique binders. If the selection process of multiple selection rounds does not result in 2 unique binders, we will start a second selection process free of charge in a 96-well plate. If this still results in less than 2 binders, we will ultimately only charge 50% of the agreed price for this step.

What is the quantity and quality of the expressed nanobody from step 2?

If the protein from step 2 is likely to be used for the initial characterization of the nanobody, 0.5 – 1 mg is often sufficient. An initial assessment of affinity, specificity and selectivity can then be made. In that case the nanobdoy is cloned into a production strain of E. coli. The protein produced is then purified via a number of steps on suitable columns and concentrated in, for example, a phosphate buffer. The purity of the nanobody from this first purification is generally >95%. For larger quantities and for other purposes the system will have to be slightly adjusted.

Which functional groups do you conjugate?

We have shown that we can conjugate multiple functional groups to nanobodies with high coupling rate, such as sCy7, NIR-800-CW, NOTA, DOTA, and DTPA, long chain fatty acids, among others. We have the most experience with maleimide conjugations to free cysteines at the C-terminus of the nanobody. For this reason, we avoid the introduction of cysteines into the CDRs of the nanobodies to prevent binding issues after conjugating functional groups. The purity of the end result can be tested with LC-MS, and is often between 90-99%.