Custom Ankyrons™

affordable rapid generation of high-quality binding reagents to new targets

 

Custom Ankyrons™ vs Custom monoclonal antibodies

Custom Ankyrons™ can be selected much faster than any monoclonal antibody can be generated through immunization and cloning.

ProImmune uses a scalable selection process allowing tens customer proteins to be screened quickly in parallel at significantly reduced cost to customers or even free of charge in the case of our privileged access program*. Custom recombinant monoclonal high specificity/affinity Ankyrons can be selected in vitro in only around 8 weeks from availability of the relevant antigen.

Unlike antibody fragments, Ankyrons do not have to undergo complex reformatting to yield functioning reagents. Expression of functional Ankyrons is not delayed long-winded processes to generate expression cell lines.

*We offer a privileged access program to free of charge custom made reagents, including raising Ankyrons to new targets in areas that are of sufficient interest to ProImmune. If you are interested in discussing this program, please contact us.

Custom Ankyrons™ feature

  • No animal immunization
  • Less time to discover than monoclonal antibodies
  • Recombinant monoclonal reagents directly selected in vitro from our Teralibrary™
  • Complicated reformatting for deployment not required
  • Rapid expression in days not weeks
  • Downstream modification including tagging and conjugation, in days not weeks
  • Guided selection can be added to the discovery process to discriminate the exact specificity you need
  • Affordable, flexible, rapid and scalable availability of follow on reagent requirements

What are Custom Ankyrons™ ideal for?

  • Rapidly obtaining recombinant monoclonal Ankyrons for individual or groups (tens) of target antigens
    • For example, imagine having rapid free or low-cost access to a complete set of binders for multiple signalling pathways or to all the proteins and their variants that are in your broader area of interest
    • Example interest areas include: Phosphorylated/non-phosphorylated proteins; Checkpoint inhibitors; therapeutic proteins and antibodies, haptenated proteins; protein-DNA complexes, MHC-peptide complexes; toxins; pesticides; environmental detection; animal research; Animal health; Crop Science; Consumer products; Research Areas: cell biology, viral proteins, bacterial proteins, neurology, autoimmunity, cardiology, immunology, PK assays, ADA, anti-idiotype binders
  • Free yourself from the many constraints of antibody technology, including time, cost, specificity, affinity, applicability, and rates of success
  • Ankyrons are suited to all the same investigational methods that antibodies are used for, such as: flow cytometry, mass cytometry, Western Blot, immunohistochemistry, ELISAs, immunofluorescence, immunoprecipitation, cell culture, ChIP, and many more
  • Perfect for guided selection between related protein targets achieving highly selective (for example against specific protein mutations/variants), or alternatively cross-reactive binding, as required
  • The easy modular design means that they can even be used as ‘molecular machines’, limited only by your imagination of how different specificities can be applied together to affect cells or cellular compartments/biochemical pathways

How to get Custom Ankyrons™

Step 1 (Requesting lab)

  • Agree with ProImmune proteins to be studied and means to provide them (are they available in the requesting lab or should ProImmune attempt to provide them)
  • Where applicable, provide proteins to ProImmune following mutual agreement on scope of proteins to be investigated
  • Ideally two nanomoles of purified proteins, e.g. 30ug for a 15kD protein or 300ug of purified IgG antibody (150kD) per protein. Where purified protein amounts are a particular problem, we can consider working with less material
  • More material may be needed for follow-up guided selection and affinity determination, where this is required
  • Where applicable, provide proteins to ProImmune following mutual agreement on scope of proteins to be investigated
  • The buffer should be amine free with near neutral pH7-8, as proteins will require amine modification for the selection process
  • Proteins should also be carrier-free, ideally with purity >85%
  • Proteins should be in a rigid folded state
  • In cases where protein targets may not be available in isolated form ProImmune may be able to generate them for the project. Please contact us for further information in that case

Step 2 (ProImmune)

  • ProImmune will aim to provide you with 4 to 6 Ankyrons per target within around 8 weeks from receipt of your target proteins
  • Ankyrons will be tested internally at ProImmune for specific binding to the target by immunoassay with >10x signal to noise ratio
  • We will typically aim to provide (where possible) at least 100ug monomeric Ankyron per clone with a standard epitope tag for secondary detection for your initial analysis and assay qualification
  • Binders are very stable and can be stored at 4°C

Step 3 (Requesting lab)

  • Test the Ankyrons in your application and identify your preferred Ankyron(s) for each target protein
  • In exchange for provision of the Ankyrons we would ask for feedback on the experimental results, including gel and staining images or flow cytometry plots where applicable with permission for ProImmune to use this information in its product literature

Step 4 (ProImmune) this will vary for each project

  • Determine accurate affinity measurements where required
  • Formatting:
    • Once basic function is confirmed we can provide additional labelled binders to most small molecule fluorophores, large fluorophores, or other commercially available labels, as well as further epitope tagged and biotinylated versions
    • Biotinylated versions can be conjugated to any streptavidin conjugate in any valency up to tetramers. Due to the typical high affinity of monomeric binders, it will not be necessary to tetramerise in all cases and a lower conjugation ratio can increase the amount of label conjugated per paratope
    • We can also provide multivalent and multimeric versions of the binders, including multi-specifics (per specific discussion/agreement with collaborators)

Step 5 (ProImmune)

  • Further assays we can perform in house include affinity determination, extended immunoassays, flow cytometry, but this may be limited by our internal capacity for such work

Contact us with your Custom Ankyron™ requirements or Request a Quote for us to generate powerful new binding reagents to your new targets quickly.