Nanobizz Services

Panning & Selection

Our synthetic libraries have a great diversity and they have their own backbone structure in which only (parts of) the CDR regions are made variable. The fixed and variable DNA fragments of these nanobodies have been cloned into plasmids that have subsequently been incorporated into suitable E. coli strains. Using M13 phages, we can then express the nanobody in question and then screen and select it in various ways. The most ideal option is to ultimately screen for targets that are identical to the intended application of the nanobody.

Cloning & Expression

After screening and selecting the desired number of different well-binding nanobodies, the desired protein can be expressed in the desired sequence using cloning. For small amounts, a few milligrams, this is the most efficient to add to E. coli. For larger quantities it is better to switch to yeast, such as Pichia pastoris.
With cloning we can introduce special amino acids that are suitable for introducing post-translational functional groups. This can be a C-terminal His-tec sequence for purification, but also a free cysteine for the induction of a fluorophore, a chelator, etc. Spacers and linkers can also be introduced with this.

Characterization

After screening and selection and any post-translational modifications, we can further characterize the selected nanobodies as desired. Determining the affinity, specificity and selectivity for the target. Initially, this is often the same cloned protein with which the screening and selection was carried out, and then tested in an ELISA or SPR setup. But it can also be extended to binding to cell fragments, whole cells or tissue.

Product Development

We can then further develop the application of a selected nanobody into, for example, a specialized ELISA, including associated validation.