Custom Affimer® service
Custom Affimer® reagents can be generated and ready for your lab in just 12-14 weeks!
Avacta’s proprietary Affimer® technology is flexible enough to meet the needs of a wide range of customers. We have undertaken numerous Affimer discovery projects for a broad variety of pharmaceutical, biotech and diagnostics companies, as well as prestigious academic institutions worldwide.
Affimer® reagents can be developed in just 12-14 weeks, compared to months for standard antibody technologies and only requires sub-milligram quantities of antigen.
The in vitro phage display selection approach, used as part of our process is not limited by the immune system and allows us to select Affimer binders with the specific desired properties for customer applications and to address difficult targets.
Affimer binders have been successfully developed for a range of applications, including:
- in vitro research
- pharmacokinetics and drug assays
- in vitro diagnostics assays
- immunoprecipitation and mass spectrometry assays
- protein purification and separation
- protein crystallisation
As part of our custom services you will get:
- Access to our proprietary libraries containing 1010 different Affimer molecules with unique properties.
- A dedicated project consultation to discuss your particular application and a flexible project design to fit your requirements.
- Regular technical interaction and reporting to maximise the chances of ideintifying the best Affimer® proteins for your needs.
- Wide range of target applicability:
- Proteins (Immunoglobulins, membrane proteins, secreted proteins, specific domains, enzymes, serum proteins, kinases)
- Small Molecules/Chemicals (e.g. TNT, diclofenac, peptidomimetics, posaconazole)
- Inorganic material (magnetic nanoparticles)
- Low target consumption – the typical process requires <100 μg of target protein
- Incoming targets QC’ed for purity, biotinylated and tested for biotinylation levels.
- Robust and well understood method using a truncated p3-fusion in the M13 phage.
- Typically 3 rounds of phage display. Selection biased in P2 and P3 to select for low off-rates.
- We routinely identify binders with <50nM Kd.
- Flexible approach in that it is possible to: Compete against related targets, vary format of target between selections, selection in buffer matched to end use, selection in presence/absence of associated ligands and screen mutant vs wild type.
Cloning and Formatting
- The phage display output is mini-prepped and the Affimer population is cloned into an expression plasmid. This allows flexibility to add tags etc. early in the process.
- Our high throughput picking and expression process allows us to assay Affimer proteins in their final format from the start of characterisation.
- No issues from assaying still on phage (multivalency, stability etc.) Nominal capacity to process ~270,000 clones per year.
Rapid Binding Screening
- Our primary screen is a bead-based high throughput iQue flow assay.
- Each clone assayed against up to 15 targets, in a fully automated 384-well plate assay.
- Protein-protein interaction inhibitor assays are available as part of the primary screen.
- This screening method offers many advantages including low target consumption (e.g. to assay 768 clones just ~7µg of target material is needed) and speed (assays can be completed in less than ½ a day).
- Primary screen data and an indication of relative clone expression levels are available by the end of week 4.
- We are currently implementing SPRi (Horiba XelPleX) and kon/koff for the clonal repertoire of each project.
- Hits from the primary screen are sequenced and unique sequences bulk expressed before moving into validation.
- Range of assays available in-house, including; fluorescence microscopy, flow cytometry, cell-based inhibition/activation assays, high-throughput pull-down assays, mass spectrometry and IHC.