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.