Services

The Core Facility Nanobodies offers both the generation of customized VHHs/nanobodies against an antigen of your choice and the production of published VHHs, for example against fluorescent proteins, common fusion partners, or specific extracellular and intracellular targets. We provide the following services:

  • Generation of custom-made VHHs
    • 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 VHH libraries that encode the VHH repertoire of a blood sample (106-107different VHHs)
    • Identification of VHHs by phage display (or advice on functional screens or alternative selection methods)
    • Identification of VHH candidates by ELISA
    • Production of a first batch of VHHs
  • Validation of custom-made VHHs
    • Binding assay in vitro
    • Binding assay in the cytosol of cultured cells
  • Generation of standard VHH expression vectors for both custom-made VHHs and VHHs with published sequences
    • for bacteria and/or mammalian cells (including lentiviral vectors)
    • as functionalized VHHs with a selection of fusion partners including fluorescent proteins, purification handles, enzymes, effector domains, etc.
  • Expression of both custom-made and published VHHs
    • different scales
    • endotoxin removal for in vivo applications
  • (Chemo-)enzymatic or chemical modification of VHHs for diverse applications. The available labeling methods include
    • Sortase A-mediated chemo-enzymatic labeling by transpeptidation
    • Amine-reactive labeling with NHS ester reactions
    • Sulfhydryl-reactive labeling with maleimide reagents

The wide range of potential applications include

  • affinity purification (VHH-coupled beads or biotinylated VHHs)
  • fluorescence microscopy (organic fluorophores such as Alexa Fluor 647®)
  • flow cytometry
  • electron microscopy (nanogold)
  • custom modifications for CyTOF, Immuno-PET, or in vivoapplications can be discussed on a case by case basis.

If you have any questions or comments on our services or custom requests not listed on our web site, please do not hesitate to contact us.