Nowadays, single-domain antibodies from camelids represent important tools in the field of biomedical research, diagnostics and therapy, which is why the phage display methodology for generating single-domain antibodies should be established at the UKS site in order to produce antibodies against antigens which cannot be marked by conventional antibodies. These so-called nanobodies are derived from heavy chain antibodies (= HcAb’s) and represent the antigen-binding part (= VHH). Compared to conventional antibodies, they convince with their small size (15 kDa), their stability against heat and pH-value changes and, with their finger-shaped paratope, they have the property to bind hidden and hard-to-reach epitopes. In addition, amplification in E. coli is fast and uncomplicated. In this work, the llama Romeo was immunized with five proteins (CaV?2, CaV?3, mFl, CAPS2, Unc119) and after confirmed immunoreaction by western blot analysis of the serum a purification of the PBL’s was performed. After RNA extraction and amplification of the VHH genes via PCR, cloning was performed into the phagemid vector pMES4, which was subsequently transformed into E. coli. The successful cloning was confirmed by colony PCR. Via superinfection with helper phages, recombinant phages, on the surface of which the nanobodies are presented, were amplified and thus a phage library was created. Subsequent panning was used to select the VHHs for the various proteins. After two to three rounds of enrichment, E. coli was infected with the selected phages and individual clones were determined and characterized in more detail. In addition to sequence analysis and subsequent classification into families (monoclonal antibodies), initial binding studies were carried out using ELISA or Western blot. After successful synthesis of specific nanobodies, single clones were given to the cooperating working groups to carry out further studies. Positive results in immunohistochemistry and X-ray crystallography are already available here. The phage display methodology described and successfully established here enables highly efficient production of single domain antibodies in E. coli cultures for use in biomedical research. Thus, experimental animals used for the production of polyclonal or monoclonal sera can be reduced in number. In addition, however, the interventions on the animal itself can be kept to a minimum due to the multiplex immunisations and the less invasive immunisation scheme, which represents a significant ethical advantage.