Standard lead development project
A full project including immunization, library construction, panning and delivery of clones can be performed within 3-4 months.
The tailor-made single-domain antibody-development projects on inquiry can include the following building blocks:
- Design and implementation of an immunization protocol in llamas
- Determination of immune responses in the sera
- Cloning of VHH-repertoires in phage-display libraries
- Panning for the desired epitope and specificity
- Picking of masterplates with monoclonal single-domain antibodies
- Production in 96 wells format
- Screening for binding in ELISA
- Sequence analysis and annotation of clusters
- Delivery of different single-domain antibodies (sequence and >0.2 mg protein).
The building blocks of the services of QVQ
We provide flexible custom projects which are tailor made for each customer based on our broad range of standard services. Services include llama immunization, construction of phage-display libraries, custom-tailored selection procedures, production of single-domain antibodies, plain or equipped with different tags, in bacteria and upscaling of production in yeast towards translation to GMP production in collaboration with Orthros TR BV.
Immunization and library construction
Immunizations of llamas are done in collaboration with Kaneka Eurogentec, Preclinics and Diaclone. Immunization strategies include the combination of several different antigens in the same animals and we can use recombinant proteins, cells, and other complex antigens. All immunizations are conducted according to the applicable animal welfare act at place and reviewed by the animal experiment commission of the company. The high quality libraries have on average a size of 10e8 different VHHs.
QVQ also offers different ready-to-go libraries, such a proprietary immune libraries, pooled immune libraries, as well as synthetic, combinatorial, or variant libraries.
Phage display selection and screening
QVQ always proposes a phage-display selection strategy that, based on own experience and available tools, should result in the selection of the desired target binders. Such a strategy is carefully designed to include customer’s requests and wishes. Importantly, the prospective final application of the single-domain antibodies are already taken into consideration while determining the phage-display selection strategy.
The expertise of the QVQ team can be used to select for agonistic or antagonistic single-domain antibodies, targeting preferred epitopes, high stability, or high specificity.
Production and delivery
After phage-display selections, individual single-domain antibodies will be characterized for binding and competition using available tools . Moreover, sequences of the different single-domain antibodies will be determined. QVQ will select a lead panel of 6 single-domain antibodies based on a combination of binding data, sequence data and, if available, functional data obtained by the customer. These lead-candidates will be produced and purified from the periplasm of Escherichia coli using IMAC.
QVQ provides a detailed report describing all the above mentioned steps and data, along with purified single-domain antibody protein of the 6 lead clones (0.2 mg of each).
Optional Follow-up Services
- NGS sequencing of outputs
- Characterization (ELISA, Flow cytometry, SPR)
- Larger scale production and purification
- Custom functionalization
- Optimization and humanization
Please contact us for more information.