Evidence for Multiple Applications of Monoclonal Antibody 5G10.
We had earlier obtained a murine monoclonal antibody (mAb), termed 5G10, that bound to Salmonella flagellin (SF) and subsequently impaired the latter property of Toll-like receptor 5 (TLR5) signaling activation. Besides interrupting SF-mediated TLR5 activation, mAb 5G10 probably had other potential applications. In this study, we explored multiple functions of 5G10. A short peptide QRVRELAV (designated T5) derived from SF in either terminal of proteins was specifically recognized by 5G10. T5 tag expressed in eukaryotic cell was also detected by 5G10 when analyzed by Western blot, immunofluorescence assay (IFA), and fluorescent-activated cell sorting (FACS). The result of the co-immunoprecipitation assay showed that 5G10 as a bait antibody dragged out the complex of enterovirus 71 (EV71) 2A and mitochondrial antiviral signaling (MAVS) protein. More importantly, 5G10 helped to purify fusion proteins T5-tagged (EV71) 2A and T5-Japanese encephalitis virus NS5 methyltransferase (MTase). Thus, it has been suggested that mAb 5G10 could be useful in several biological applications, including protein identification, location, and affinity purification.
['Xie|Lilan|L|', 'Chen|Wangsheng|W|', 'Li|Jianjun|J|', 'Li|Yi|Y|', 'Li|Yaoming|Y|']
D048868:Adaptor Proteins, Signal Transducing D000818:Animals D000911:Antibodies, Monoclonal D002846:Chromatography, Affinity D004664:Encephalitis Virus, Japanese D005408:Flagellin D006801:Humans D051379:Mice D010455:Peptides D011485:Protein Binding D011993:Recombinant Fusion Proteins D012475:Salmonella D015398:Signal Transduction D051198:Toll-Like Receptor 5 D017361:Viral Nonstructural Proteins
monoclonal antibody flagellin protein purification