@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}
}