@misc{HanackSchloerHolzloehneretal.2016,
  author    = {Hanack, Katja and Schloer, Anja and Holzloehner, Pamela and Listek, Martin and Bauer, Cindy and Butze, Monique and Micheel, Burkhard and Hentschel, Christian and Sowa, Mandy and Roggenbuck, Dirk and Schierack, Peter and Fuener, Jonas and Schliebs, Erik and Goihl, Alexander and Reinhold, Dirk},
  title     = {Camelid nanobodies specific to human pancreatic glycoprotein 2},
  series = {The journal of immunology},
  volume    = {196},
  journal   = {The journal of immunology},
  publisher = {American Assoc. of Immunologists},
  address   = {Bethesda},
  issn      = {0022-1767},
  pages     = {313 -- 328},
  year      = {2016},
  abstract  = {Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified to be a major autoantigenic target in Crohn's disease patients. It was discussed recently that a long and a short isoform of GP2 exists whereas the short isoform is often detected by GP2-specific autoantibodies. In the outcome of inflammatory bowel diseases, these GP2-specific autoantibodies are discussed as new serological markers for diagnosis and therapeutic monitoring. To investigate this further, camelid nanobodies were generated by phage display and selected against the short isoform of GP2 in order to isolate specific tools for the discrimination of both isoforms. Nanobodies are single domain antibodies derived from camelid heavy chain only antibodies and characterized by a high stability and solubility. The selected candidates were expressed, purified and validated regarding their binding properties in different enzyme-linked immunosorbent assays formats, immunofluorescence, immunohistochemistry and surface plasmon resonance spectroscopy. Four different nanobodies could be selected whereof three recognize the short isoform of GP2 very specifically and one nanobody showed a high binding capacity for both isoforms. The KD values measured for all nanobodies were between 1.3 nM and 2.3 pM indicating highly specific binders suitable for the application as diagnostic tool in inflammatory bowel disease.},
  language  = {en}
}
@misc{MaierHolzloehnerHoenowetal.2016,
  author    = {Maier, Natalia and Holzl{\"o}hner, Pamela and Hoenow, Anja and Scheunemann, Astrid and Weschke, Daniel and Hanack, Katja},
  title     = {Characterization of monoclonal antibodies generated by in vitro immunization},
  series = {The journal of immunology},
  volume    = {196},
  journal   = {The journal of immunology},
  publisher = {American Assoc. of Immunologists},
  address   = {Bethesda},
  issn      = {0022-1767},
  pages     = {2},
  year      = {2016},
  abstract  = {Monoclonal antibodies are highly valuable tools in biomedicine but the generation by hybridoma technology is very time-consuming and elaborate. In order to circumvent the consisting drawbacks an in vitro immunization approach was established by which murine as well as human monoclonal antibodies against a viral coat protein could be developed. The in vitro immunization process was performed by isolation of murine hematopoietic stem cells or human monocytes and an in vitro differentiation into immature dendritic cells. After antigen loading the cells were co-cultivated with naive T and B lymphocytes for three days in order to obtain antigen-specific B lymphocytes in culture, followed by fusion with murine myeloma cells or human/murine heteromyeloma cells. Antigen-specific hybridomas were selected and the generated antibodies were purified and characterized in this study by ELISA, western blot, gene sequencing, affinity measurements. Further the characteristics were compared to a monoclonal antibody against the same target generated by conventional hybridoma technology. Isotype detection revealed a murine IgM and a human IgG4 antibody in comparison to an IgG1 for the conventionally generated antibody. The antibodies derived from in vitro immunization showed indeed a lower affinity for the antigen as compared to the conventionally generated one, which is probably based on the significantly shorter B cell maturation (3 days) during the immunization process. Nevertheless, they were suitable for building up a sandwich based detection system. Therefore, the in vitro immunization approach seems to be a good and particularly fast alternative to conventional hybridoma technology.},
  language  = {en}
}
@misc{HolzloehnerButzeHebeletal.2016,
  author    = {Holzl{\"o}hner, Pamela and Butze, Monique and Hebel, Nicole and Weschke, Daniel and Schliebs, E. and Naumann, F. and F{\"u}ner, J. and Micheel, Burkhard and Hanack, Katja},
  title     = {Monoclonal mouse antibodies against PBMC subpopulations of New World camelides},
  series = {European journal of immunology},
  volume    = {46},
  journal   = {European journal of immunology},
  publisher = {Wiley-Blackwell},
  address   = {Hoboken},
  issn      = {0014-2980},
  pages     = {1175 -- 1175},
  year      = {2016},
  language  = {en}
}