@Article{ecn.2018.0409,
AUTHOR = {Karinna Chouman, Birgit Korioth-Schmitz, Markus Sack, Jörn Engelbert Schmitz, Anh Tuan Pham, Rainer Fischer, Stefan Barth, Torsten Klockenbring, Rolf Fendel},
TITLE = {Characterization of new anti-IL-6 antibodies revealed high potency candidates for intracellular cytokine detection and specific targeting of IL-6 receptor binding sites},
JOURNAL = {European Cytokine Network},
VOLUME = {29},
YEAR = {2018},
NUMBER = {2},
PAGES = {59--72},
URL = {http://www.techscience.com/ECN/v29n2/65406},
ISSN = {1952-4005},
ABSTRACT = {Interleukin-6 (IL-6) expression and secretion, induced by inflammatory processes, stimulate the acute
phase response cascade. The overexpression of IL-6 contributes to a variety of inflammatory diseases, <i>e.g.</i> rheumatoid
arthritis, Castleman’s disease, multiple myeloma, and prostate cancer. Screening for high amounts of IL-6 in
the patients’ blood serum can be crucial for an adequate treatment. In this study, five novel murine monoclonal
antibodies (mAbs) reactive to human IL-6 were generated. The mAbs were characterized for potential diagnostic
purposes and recombinant antibodies were derived thereof. Initial epitope mapping using a combination of blocking
experiments and Hyper-IL-6, a fusion protein consisting of IL-6 and the soluble IL-6 receptor revealed distinct but
overlapping binding sites. At least one of the mAbs was found to interact with the region of IL-6/ IL-R complex
formation. Three mAbs were applied successfully in intracellular staining by flow cytometry, whereas one of the
mAbs showed comparable binding as a reference reagent. Furthermore, the mAbs were tested for applications in
various immunological assays such as ELISA, Western blot and surface plasmon resonance spectroscopy (SPR),
using IL-6 from commercial sources as well as in-house produced protein (IL-6_IME). The limit of detection was
determined by sandwich ELISA (0.5 ng/mL, SD ±0.005). Our results also demonstrated that the recombinant IL-6
produced was functional and correctly folded. These findings support the use of the generated mAb clones as
promising candidates for application in various immunological assays for diagnostic and scientific purposes.},
DOI = {10.1684/ecn.2018.0409}
}