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.

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.