Antibodies
Prof. Dr. Ilse Hofmann
Group Leader
Welcome to the Core Facility Antibodies. We offer the production and purification of antibodies. These include: monoclonal antibodies (hybridoma technology, mouse/rat), polyclonal ABs (guinea pig) and recombinant antibodies (sequencing, engineering). Many of the antibodies produced in-house are available for internal research purposes (antibody list).
Services
monoclonal antibodies (hybridoma) | polyclonal antibodies | recombinant antibodies | purification | measurement of biomolecular interactions
Services - Information
We use state-of-the-art hybridoma technologies for production of monoclonal antibodies via the somatic fusion of lymphocytes from immunized mice or rats. We advise in the selection of antigens, immunize animals and perform screening of mother clones by e.g. ELISA, immunofluorescence or IHC. Clones having been tested successfully will be subcloned, expanded and maintained to guarantee long-time availability of materials. In addition, we provide non-GMP production and purification of antibodies.
Our service includes:
- Advice in the selection of the antigen (protein/peptide)
- Alignment analysis to check for cross-reactivity
- Immunization of animals
- Titer test
- Advice in validation of antibody specificity
- Consultation and close interaction with customers during all steps of production
Our workflow:
- Fusion of lymphocytes to immortalized cell line
- Screening of clones by ELISA/Western blot/Immunofluorescence
- Selection of clones displaying desired specificity
- Sub-cloning to capture stable cell lines
- Estimation of antibody subgroup
- Expansion and storage of positive clones
- Supply with supernatant for your experiments
- If required, scale up and antibody purification
Polyclonal antibody production (guinea pig)
The CF Antibodies offers the production of polyclonal (guinea pig) ABs, that specifically recognize antigens giving low antibody responses in mice or rats.
Our service includes:
- Advice in the selection of the antigen (protein/peptide)
- Alignment analysis to check for cross-reactivity
- Immunisation of animals with various approaches
- Titer test
- Advice in validation of antibody specificity
You supply:
- Peptides conjugated to KLH for immunization and unconjugated for ELISA screening
- Recombinant protein preferable with HIS-tag; we can support you in the generation of HIS-tagged proteins.
Recombinant antibodies
For recombinant antibody production in HEK cells, RNA of hybridomas is sequenced, the variable immunoglobulin regions identified, cloned in frame into different Fc backbones and the immunoglobulin chains are expressed.
- RNA isolation and RNA sequencing of stable hybridoma cell line
- Identification of variable regions of the heavy and light chain
- Cloning into Fc backbone (human, mouse, rabbit, Fab with HIS- or Streptavidin-tag), scFv
- Antibody production through transient expression in HEK cells
Many of our antibodies are available for internal research at the DKFZ (hybridoma supernatant or purified).
These are mostly proprietary antibodies that are not readily available from other sources.
List of available antibodies
(opens website on other DKFZ server, only available in English)
Measurement of biomolecular interactions
Microscale Thermophoresis MST
- Measurement in glass capillaries
- low sample requirement
- Robust and easy handling
- Determination of binding affinities
We offer access to the device, guidance and support (“assisted access”)
For further information Microscale Thermophoresis MST
(opens website on other DKFZ server, only available in English)
Team
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Prof. Dr. Ilse Hofmann
Head of Unit
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Tanja Schlechter
Technician
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Natalie Erbe-Hofmann
Technician
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Claudia Linhard
Technician
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Fabian Zimmermann
Technician
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Dr. Frank Schwarz
Biochemist & Computer Scientist
Selected Publications
Kim M., Reidenbach S., Schlechter T., Rothmann AC., Will R., Hofmann I. Plakophilin 1-deficient cells up-regulate cytokine expression and this correlates with immune cell recruitment in prostatic tumors. (2023) FEBS Apr.290(7)1907-1919 PMID: 36380691
Nikitina E, Burk-Körner A, Wiesenfarth M, Alwers E, Heide D, Tessmer C, Ernst C, Krunic D, Schrotz-King P, Chang-Claude J, von Winterfeld M, Herpel E, Brobeil A, Brenner H, Heikenwalder M, Hoffmeister M, Kopp-Schneider A, Bund T. Bovine meat and milk factor protein expression in tumor-free mucosa of colorectal cancer patients coincides with macrophages and might interfere with patient survival. (2023) Mol Oncol. Feb 22. PMID: 36811271 doi: 10.1002/1878-0261.13390
Phillips E., Balss J., Bethke F., Pusch S., Christen S., Hielscher T., Schnölzer M., Fletcher MNC., Habel A., Tessmer C., Brenner LM., Göttmann M., Capper D., Herold-Mende C., von Deimling A., Fendt SM., Goidts V. PFKFB4 interacts with FBXO28 to promote HIF-1α signaling in glioblastoma. Oncogenesis. 2022 Sep 17;11(1):57 PMID: 36115843
Sicong Ma, Roger Sandhoff, Xiu Luo, Fuwei Shang, Qiaozhen Shi, Zhaolong Li, Jingxia Wu, Yanan Ming, Frank Schwarz, Alaa Madi, Nina Weisshaar, Alessa Mieg, Marvin Hering, Ferdinand Zettl, Xin Yan, Kerstin Mohr, Nora Ten Bosch, Zhe Li, Gernot Poschet, Hans-Reimer Rodewald, Nina Papavasiliou, Xi Wang, Pu Gao, Guoliang Cui
Serine enrichment in tumors promotes regulatory T cell accumulation through sphinganine-mediated regulation of c-Fos
Sci Immunol. 2024 Apr 19;9(94):eadg8817. doi: 10.1126/sciimmunol.adg8817. Epub 2024 Apr 19. PMID:38640251
Pfefferle S , Schöpf J, Kögl M, Friedel C, Müller M, Carbajo-Lozoya J, Stellberger T, von Dall'Armi E, Herzog P, Kallies S, Niemeyer D, Vanessa Ditt V, Kuri T, Züst R, Pumpor K, Hilgenfeld R, Schwarz F, Zimmer R, Steffen I, Weber F, Thiel V, Herrler G, Thiel HJ, Schwegmann-Wessels C, Pöhlmann S, Haas J, Drosten Christian, von Brunn A
The SARS-coronavirus-host interactome: identification of cyclophilins as target for pan-coronavirus inhibitors
PLoS Pathog. 2011 Oct;7(10):e1002331. Epub 2011 Oct 27 PMID: 22046132