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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 intervals. To maximise chances of raising a good immune response against most injected targets, we practice immunisation with 4 to 5 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 antibodies).
Animal ordinance requires that samples for immunisation are free from toxic and irritating substances (DTT, urea, guanidine hydrochloride, sodium azide, and similar).
Following immunisation, approximately 80 ml blood is collected to obtain lymphocyte mRNA. The genetic material is then cloned into a phage display vector. Routinely, phage libraries greater than 1x108 individual clones are obtained and screened in consecutive rounds of binding, elution and amplification of phages which specifically bind to the target. Usually 2-3 rounds of panning are necessary to sufficiently enrich the phages of interest. The final validation of binders is done by ELISA screening of 190 clones per target.
Subsequent characterization of variability of identified binders from the nano-antibody library is done by sequencing 96 ELISA-positive clones.
The nano-antibodies of interest are then expressed recombinantly using different expression systems and purified using affinity chromatography by the final users and can be further characterized in analytical and functional assay of choice.