Functional knockdown of VCAM-1 at the posttranslational level with ER retained antibodies
Vascular cell adhesion molecule 1 (VCAM-1) is involved in the recruitment of leukocytes to inflammatory sites. In this study we present the first functional knockdown of VCAM-1 using an ER retained antibody construct. We generated a knockdown construct encoding the VCAM-1 specific single chain varia...
Gespeichert in:
| Hauptverfasser: | , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
25 November 2008
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| In: |
Journal of immunological methods
Year: 2009, Jahrgang: 341, Heft: 1/2, Pages: 30-40 |
| ISSN: | 1872-7905 |
| DOI: | 10.1016/j.jim.2008.10.012 |
| Online-Zugang: | Verlag, Volltext: http://dx.doi.org/10.1016/j.jim.2008.10.012 Verlag, Volltext: http://www.sciencedirect.com/science/article/pii/S0022175908003232 
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| Verfasserangaben: | N. Strebe, A. Guse, M. Schüngel, T. Schirrmann, M. Hafner, T. Jostock, M. Hust, W. Müller, S. Dübel |
MARC
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| 520 | |a Vascular cell adhesion molecule 1 (VCAM-1) is involved in the recruitment of leukocytes to inflammatory sites. In this study we present the first functional knockdown of VCAM-1 using an ER retained antibody construct. We generated a knockdown construct encoding the VCAM-1 specific single chain variable fragment scFv6C7.1 fused to the C-terminal ER retention sequence KDEL. HEK-293:VCAM-YFP cells stably expressing a VCAM-YFP fusion protein were transiently transfected with the knockdown construct and showed down-regulation of surface VCAM-1. Knockdown efficiency of the system is time-dependent due to used transient transfection of the intrabody construct. Furthermore, intrabody mediated knockdown of HEK-293:VCAM-YFP cells also impaired cell-cell interaction with Jurkat cells that are endogenously expressing VLA-4, the physiological partner of VCAM-1. Posttranslational knockdown with ER retained antibodies seems to be a promising technique, as shown here for VCAM-1. | ||
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| 650 | 4 | |a Intrabody | |
| 650 | 4 | |a Knockdown | |
| 650 | 4 | |a Recombinant antibody | |
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| 650 | 4 | |a VCAM-1 | |
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