@misc{GoethelListekMesserschmidtetal.2021,
  author    = {G{\"o}thel, Markus and Listek, Martin and Messerschmidt, Katrin and Schl{\"o}r, Anja and H{\"o}now, Anja and Hanack, Katja},
  title     = {A New Workflow to Generate Monoclonal Antibodies against Microorganisms},
  series = {Mathematisch-Naturwissenschaftliche Reihe},
  journal   = {Mathematisch-Naturwissenschaftliche Reihe},
  number    = {20},
  issn      = {1866-8372},
  doi       = {10.25932/publishup-52334},
  url       = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-523341},
  pages     = {17},
  year      = {2021},
  abstract  = {Monoclonal antibodies are used worldwide as highly potent and efficient detection reagents for research and diagnostic applications. Nevertheless, the specific targeting of complex antigens such as whole microorganisms remains a challenge. To provide a comprehensive workflow, we combined bioinformatic analyses with novel immunization and selection tools to design monoclonal antibodies for the detection of whole microorganisms. In our initial study, we used the human pathogenic strain E. coli O157:H7 as a model target and identified 53 potential protein candidates by using reverse vaccinology methodology. Five different peptide epitopes were selected for immunization using epitope-engineered viral proteins. The identification of antibody-producing hybridomas was performed by using a novel screening technology based on transgenic fusion cell lines. Using an artificial cell surface receptor expressed by all hybridomas, the desired antigen-specific cells can be sorted fast and efficiently out of the fusion cell pool. Selected antibody candidates were characterized and showed strong binding to the target strain E. coli O157:H7 with minor or no cross-reactivity to other relevant microorganisms such as Legionella pneumophila and Bacillus ssp. This approach could be useful as a highly efficient workflow for the generation of antibodies against microorganisms.},
  language  = {en}
}
@article{GoethelListekMesserschmidtetal.2021,
  author    = {G{\"o}thel, Markus and Listek, Martin and Messerschmidt, Katrin and Schl{\"o}r, Anja and H{\"o}now, Anja and Hanack, Katja},
  title     = {A New Workflow to Generate Monoclonal Antibodies against Microorganisms},
  series = {Applied Sciences},
  volume    = {11},
  journal   = {Applied Sciences},
  number    = {20},
  publisher = {MDPI},
  address   = {Basel},
  issn      = {1454-5101},
  doi       = {10.3390/app11209359},
  pages     = {15},
  year      = {2021},
  abstract  = {Monoclonal antibodies are used worldwide as highly potent and efficient detection reagents for research and diagnostic applications. Nevertheless, the specific targeting of complex antigens such as whole microorganisms remains a challenge. To provide a comprehensive workflow, we combined bioinformatic analyses with novel immunization and selection tools to design monoclonal antibodies for the detection of whole microorganisms. In our initial study, we used the human pathogenic strain E. coli O157:H7 as a model target and identified 53 potential protein candidates by using reverse vaccinology methodology. Five different peptide epitopes were selected for immunization using epitope-engineered viral proteins. The identification of antibody-producing hybridomas was performed by using a novel screening technology based on transgenic fusion cell lines. Using an artificial cell surface receptor expressed by all hybridomas, the desired antigen-specific cells can be sorted fast and efficiently out of the fusion cell pool. Selected antibody candidates were characterized and showed strong binding to the target strain E. coli O157:H7 with minor or no cross-reactivity to other relevant microorganisms such as Legionella pneumophila and Bacillus ssp. This approach could be useful as a highly efficient workflow for the generation of antibodies against microorganisms.},
  language  = {en}
}
@misc{ListekHoenowGossenetal.2020,
  author    = {Listek, Martin and H{\"o}now, Anja and Gossen, Manfred and Hanack, Katja},
  title     = {A novel selection strategy for antibody producing hybridoma cells based on a new transgenic fusion cell line},
  series = {Postprints der Universit{\"a}t Potsdam : Mathematisch Naturwissenschaftliche Reihe},
  journal   = {Postprints der Universit{\"a}t Potsdam : Mathematisch Naturwissenschaftliche Reihe},
  number    = {865},
  issn      = {1866-8372},
  doi       = {10.25932/publishup-45989},
  url       = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-459893},
  pages     = {14},
  year      = {2020},
  abstract  = {The use of monoclonal antibodies is ubiquitous in science and biomedicine but the generation and validation process of antibodies is nevertheless complicated and time-consuming. To address these issues we developed a novel selective technology based on an artificial cell surface construct by which secreted antibodies were connected to the corresponding hybridoma cell when they possess the desired antigen-specificity. Further the system enables the selection of desired isotypes and the screening for potential cross-reactivities in the same context. For the design of the construct we combined the transmembrane domain of the EGF-receptor with a hemagglutinin epitope and a biotin acceptor peptide and performed a transposon-mediated transfection of myeloma cell lines. The stably transfected myeloma cell line was used for the generation of hybridoma cells and an antigen- and isotype-specific screening method was established. The system has been validated for globular protein antigens as well as for haptens and enables a fast and early stage selection and validation of monoclonal antibodies in one step.},
  language  = {en}
}
@article{ListekHoenowGossenetal.2020,
  author    = {Listek, Martin and H{\"o}now, Anja and Gossen, Manfred and Hanack, Katja},
  title     = {A novel selection strategy for antibody producing hybridoma cells based on a new transgenic fusion cell line},
  series = {Scientific Reports},
  volume    = {10},
  journal   = {Scientific Reports},
  publisher = {Macmillan Publishers Limited, part of Springer Nature},
  address   = {London},
  issn      = {2045-2322},
  doi       = {10.1038/s41598-020-58571-w},
  pages     = {12},
  year      = {2020},
  abstract  = {The use of monoclonal antibodies is ubiquitous in science and biomedicine but the generation and validation process of antibodies is nevertheless complicated and time-consuming. To address these issues we developed a novel selective technology based on an artificial cell surface construct by which secreted antibodies were connected to the corresponding hybridoma cell when they possess the desired antigen-specificity. Further the system enables the selection of desired isotypes and the screening for potential cross-reactivities in the same context. For the design of the construct we combined the transmembrane domain of the EGF-receptor with a hemagglutinin epitope and a biotin acceptor peptide and performed a transposon-mediated transfection of myeloma cell lines. The stably transfected myeloma cell line was used for the generation of hybridoma cells and an antigen- and isotype-specific screening method was established. The system has been validated for globular protein antigens as well as for haptens and enables a fast and early stage selection and validation of monoclonal antibodies in one step.},
  language  = {en}
}
@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}
}
@article{SchloerHolzloehnerListeketal.2018,
  author    = {Schl{\"o}r, Anja and Holzl{\"o}hner, Pamela and Listek, Martin and Grieß, Cindy and Butze, Monique and Micheel, Burkhard and Hentschel, Christian and Sowa, Mandy and Roggenbuck, Dirk and Schierack, Peter and F{\"u}ner, Jonas and Schliebs, Erik and Goihl, Alexander and Reinhold, Dirk and Hanack, Katja},
  title     = {Generation and validation of murine monoclonal and camelid recombinant single domain antibodies specific for human pancreatic glycoprotein 2},
  series = {New biotechnology},
  volume    = {45},
  journal   = {New biotechnology},
  publisher = {Elsevier},
  address   = {Amsterdam},
  issn      = {1871-6784},
  doi       = {10.1016/j.nbt.2018.03.006},
  pages     = {60 -- 68},
  year      = {2018},
  abstract  = {Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified as a major autoantigenic target in Crohn's disease patients. It was reported recently that a long (GP2a) and a short (GP2b) isoform of GP2 exist and that in the outcome of inflammatory bowel diseases (IBD) GP2-specific autoantibodies probably appear as new serological markers for diagnosis and therapeutic monitoring. To investigate this further and in order to establish diagnostic tools for the discrimination of both GP2 isoforms, a set of different murine monoclonal and camelid recombinant single domain antibodies (camelid VHH) was generated and validated in various enzyme-linked immunosorbent assay (ELISA) formats, immunofluorescence on transgenic cell lines and immunohistochemistry on monkey pancreas tissue sections. Out of six binders identified, one was validated as highly specific for GP2a. This murine monoclonal antibody (mAb) was used as capture antibody in construction of a sandwich ELISA for the detection of GP2a. Camelid VHHs or a second murine mAb served as detection antibodies in this system. All antibodies were also able to stain GP2a or GP2b on transgenic cell lines as well as on pancreatic tissue in immunohistochemistry. The KD values measured for the camelid VHHs were between 7 nM and 23pM. This set of specific binders will enable the development of suitable diagnostic tools for GP2-related studies in IBD.},
  language  = {en}
}
@inproceedings{ListekMicheelHeilmann2012,
  author    = {Listek, Martin and Micheel, Burkhard and Heilmann, K.},
  title     = {Insertion of artificial cell surface receptors for antigen-specific labelling of hybridoma cells},
  series = {Immunology : an official journal of the British Society for Immunology},
  volume    = {137},
  booktitle = {Immunology : an official journal of the British Society for Immunology},
  publisher = {Wiley-Blackwell},
  address   = {Hoboken},
  issn      = {0019-2805},
  pages     = {651 -- 651},
  year      = {2012},
  language  = {en}
}
@article{FudickarRoderListeketal.2021,
  author    = {Fudickar, Werner and Roder, Phillip and Listek, Martin and Hanack, Katja and Linker, Torsten},
  title     = {Pyridinium alkynylanthracenes as sensitizers for photodynamic therapy},
  series = {Photochemistry and photobiology},
  volume    = {98},
  journal   = {Photochemistry and photobiology},
  number    = {1},
  publisher = {Wiley},
  address   = {Hoboken},
  issn      = {0031-8655},
  doi       = {10.1111/php.13554},
  pages     = {193 -- 201},
  year      = {2021},
  abstract  = {Photodynamic therapy (PDT) is a mild but effective method to treat certain types of cancer upon irradiation with visible light. Here, three isomeric methylpyridinium alkynylanthracenes 1op were evaluated as sensitizers for PDT. Upon irradiation with blue or green light, all three compounds show the ability to initiate strand breaks of plasmid DNA. The mayor species responsible for cleavage is singlet oxygen (O-1(2)) as confirmed by scavenging reagents. Only isomers 1m and 1p can be incorporated into HeLa cells, whereas isomer 1o cannot permeate through the membrane. While isomer 1m targets the cell nucleus, isomer 1p assembles in the cellular cytoplasm and impacts the cellular integrity. This is in accordance with a moderate toxicity of 1p in the dark, whereas 1m exhibits no dark toxicity. Both isomers are suitable as PDT reagents, with a CC50 of 3 mu m and 75 nm, for 1p and 1m, respectively. Thus, derivative 1m, which can be easily synthesized, becomes an interesting candidate for cancer therapy.},
  language  = {en}
}