The Core Facility Nanobodies offers the generation of customized VHHs/nanobodies using the following procedures:

  • Immunization of alpacas with the purified target protein of choice (other types of immunogens can be discussed on a case by case basis)
  • Testing alpaca sera for antibodies against the immunogen
  • Generation of nanobody libraries that encode the VHH repertoire of a blood sample (106-107 different VHHs)
  • Identification of VHHs by phage display (or advice on functional screens or alternative selection methods)
  • Identification of VHH candidates by ELISA
  • Small scale production and validation of VHHs (binding tested in vitro and in the cytosol of cells as needed)
  • Generation of nanobody expression vectors for bacteria and mammalian cells (including lentiviral vectors) – VHHs can be functionalized with a selection of fusion partners including fluorophores, purification handles, and enzymes
  • Production of a first batch of VHH
  • Enzymatic or chemical modification of VHHs for applications in
    1. Affinity purification (VHH-coupled beads or biotinylated VHHs)
    2. Fluorescence microscopy or flow cytometry (organic fluorophores such as Alexa Fluor 647®)
    3. Electron microscopy (nanogold)
    4. Further modifications for CyTOF, Immuno-PET, or in vivo applications can be discussed on a case by case basis

We also produce and enzymatically label published nanobodies (e.g. against fluorescent proteins, common fusion partners, or specific extracellular and intracellular targets).