The DiViNe partners will use a new generation of affinity chromatography ligands, Nanofitins®, with capture features being at least similar to Protein A when used to purified IgG, but whose applications could extend far beyond the sole field of antibodies

Performance comparable with commercial productsNanofitin ligands 1

Nanofitins® allow capture and elution of selected macromolecules ranging from small peptides to viruses. Nanofitins® have already been tested in comparison with Protein A with a level of performance comparable to most available commercial products. They could exhibit the same level of performance against virtually any macromolecular target in affinity columns or membranes. In order to enhance the capture capacity of the resins, it is possible to genetically engineer Nanofitins®. Nanofitins® are also particularly amenable to immunoprecipitation processes thanks to their robustness.

Custom purification with tailored elution parameters

Usual selection process is a 2 months effort and yields to a number of different monomeric hits with a high affinity down to single-digit picomolar. However, affinity can be tailored in the 10E-5 – 10E-12 range directly from within the selection process for affinity chromatography needs where release of the target is an essential step. However, high affinity Nanofitins® will also be retained in the DiViNe project with a minor effort for subsequent use in QC assays (ELISA, Western Blot,…) as they represent a promising alternative to antibodies for immunoassays as well.

In the case of Protein A capture of antibodies, the usual elution is conducted through a drastic pH shift (pH = 2.5). This can extremely damaging for some products. The Nanofitin® discovery process allows choosing and engineering ligands that can dissociate from the target under previously selected conditions.

Straightforward conjugation process

Engineering NanofitinsThe conserved region, common to all Nanofitins®, bears the regions used for conjugation to resins, beads, membranes,… In particular, regioselective conjugation is possible through insertion of one single Cysteine in the Nanofitin® sequence, thereby offering identical thiol-based covalent coupling. All Nanofitins® can undergo the very same process for incorporation into chromatography media, with similar behavior in terms of association to the material.

Promising safety profile

Target binding by Nanofitins® is also applied by Affilogic to provide in vivo neutralizing agents as therapeutics. While the field is totally different, outputs from each therapeutic program, notably in terms of safety and stability of Nanofitins® provide very promising and useful data and technologies for a safe purification process.