Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7

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JAKOBS, Barbara D., Lisa SPANNAGEL, Vladimir PURVANOV, Edith UETZ-VON ALLMEN, Christoph MATTI, Daniel F. LEGLER, 2019. Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7. In: International Journal of Molecular Sciences. 20(10), 2597. eISSN 1422-0067. Available under: doi: 10.3390/ijms20102597

@article{Jakobs2019-05-27Engin-46455, title={Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7}, year={2019}, doi={10.3390/ijms20102597}, number={10}, volume={20}, journal={International Journal of Molecular Sciences}, author={Jakobs, Barbara D. and Spannagel, Lisa and Purvanov, Vladimir and Uetz-von Allmen, Edith and Matti, Christoph and Legler, Daniel F.}, note={Article Number: 2597} }

Legler, Daniel F. Uetz-von Allmen, Edith Purvanov, Vladimir eng Purvanov, Vladimir Matti, Christoph Uetz-von Allmen, Edith 2019-07-18T12:07:31Z Jakobs, Barbara D. 2019-07-18T12:07:31Z Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7 Jakobs, Barbara D. Matti, Christoph The chemokine receptor CCR7 plays a pivotal role in health and disease. In particular, CCR7 controls homing of antigen-bearing dendritic cells and T cells to lymph nodes, where adaptive immune responses are initiated. However, CCR7 also guides T cells to inflamed synovium and thereby contributes to rheumatoid arthritis and promotes cancer cell migration and metastasis formation. Nanobodies have recently emerged as versatile tools to study G-protein-coupled receptor functions and are being tested in diagnostics and therapeutics. In this study, we designed a strategy to engineer novel nanobodies recognizing human CCR7. We generated a nanobody library based on a solved crystal structure of the nanobody Nb80 recognizing the β<sub>2</sub>-adrenergic receptor (β<sub>2</sub>AR) and by specifically randomizing two segments within complementarity determining region 1 (CDR1) and CDR3 of Nb80 known to interact with β<sub>2</sub>AR. We fused the nanobody library to one half of split-YFP in order to identify individual nanobody clones interacting with CCR7 fused to the other half of split-YFP using bimolecular fluorescence complementation. We present three novel nanobodies, termed Nb1, Nb5, and Nb38, that recognize human CCR7 without interfering with G-protein-coupling and downstream signaling. Moreover, we were able to follow CCR7 trafficking upon CCL19 triggering using Nb1, Nb5, and Nb38. Legler, Daniel F. Attribution 4.0 International Spannagel, Lisa Spannagel, Lisa 2019-05-27

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