Service and fees

We provide the full range of services starting from library production and binder selection to characterization of target-specific binders.

Our immunisation strategy includes 4 injections of antigen administered in a two week interval. To maximise chances of raising a good immune response against all injected targets, we practice immunisation with maximum of 4 purified antigens per animal. The success of raising a specific immune response depends greatly on the nature and the quality of the target antigen and it is serologically monitored for the increasing titre of IgG2 and IgG3 subclass (heavy-chain-only antibody).

Animal ordinance requires that samples for immunisation are free from toxic and irritating substances (DTT, urea, guanidine hydrochloride, sodium azide, etc.).

Following immunisation, approximately 60-80 ml blood is collected to obtain lymphocyte mRNA. The genetic material is then cloned into a phage display vector. Routinely, phage libraries of 1x10⁷ - 1x10⁸ individual colonies are obtained and screened in consecutive rounds of binding, elution and amplification of phages which specifically bind to the target. Usually 2-4 rounds of panning are necessary to sufficiently enrich the phages of interest. This phase requires one month time and is succeeded by ELISA screening of at least two microtiter plates (192 colonies) per target.

Subsequent characterization of variability of identified binders from the nanobody library is done by sequencing of ELISA-positive clones.

The nanobodies of interest can be expressed recombinantly using different expression systems and purified using affinity chromatography.

Nanobodies can be further characterized in analytical and functional assay of choice.