The Core Facility Nanobodies offers both the generation of customized nanobodies against an antigen of your choice and the production of nanobodies from known sequences. We provide the following services:
- Generation of custom-made nanobodies
- Immunization of a llama or alpaca with the target antigen (usually purified protein provided by the user, other immunization strategies can be discussed)
- Serum ELISA to verify immune response against the target antigen
- Generation of VHH phage libraries encoding the VHH repertoire of a blood sample (106-107different VHHs)
- Enrichment of target specific VHHs by phage display (different selection strategies)
- Identification of nanobody candidates by ELISA
- Small scale production and purification of preliminary hits
- Validation of custom-made nanobodies
- Binding assay in vitro
- Binding assay in the cytosol of cultured cells
- Generation of standard nanobody expression vectors for both custom-made nanobodies and nanobodies with published sequences
- For expression of monovalent nanobodies in bacteria and/or mammalian cells (including lentiviral vectors)
- For expression of functionalized nanobodies with a fusion partner (e.g. fluorescent proteins, purification tags, enzymes, etc.)
- For expression different nanobody formats formats including dimers and Fc fusion proteins
- Expression of both custom-made and published nanobodies
- Small scale / large scale
- endotoxin removal for in vivoapplications
- Enzymatic or chemical modification of VHHs for diverse applications.
- Sortase A-mediated chemo-enzymatic labeling by transpeptidation (to site-specifically attach fluorophores, oligonucleotides, biotin etc.)
- Amine-reactive labeling with NHS ester reactions (to install biotin or fluorophores on exposed lysines or prepare VHH-coupled magnetic beads)
Popular nanobody applications (performed by customers)
- fluorescence microscopy (organic fluorophores such as Alexa Fluor 488/647®)
- flow cytometry (fluorophores or secondary staining)
- structural biology (crystallization, cryo-EM)
- affinity purification (nanobodies coupled to beads or biotin)
- in vivo imaging (fluorophores, PET emitters)
- functional modulation of target proteins (blocking or activating)
- modulation of protein interactions (induction or inhibition)
- therapeutic applications (enzyme inhibition, modulation of protein-protein interaction, cell depletion, bispecifics, CARs, BiTEs, ADCs)
Please don´t hesitate to contact us. We are looking forward to discuss all your questions and ideas.