• Raffi Manoukian, Hong Sun, Silke Miller, Di Shi, Brian Chan, and Cen Xu.
• Department of Cytometry Sciences, Amgen Research, 360 Binney Street, Cambridge, MA, 02142, USA.
• J Headache Pain. 2019 Apr 30; 20 (1): 44.

BackgroundMonoclonal antibodies against calcitonin gene-related peptide (CGRP) or its receptor are efficacious for the prevention of migraine headaches. The downstream molecular mechanisms following ligand-receptor blockade by which these antibodies prevent CGRP signaling through CGRP receptors have not been demonstrated.MethodsHere we produced tool monoclonal functional antagonist antibodies against CGRP and its canonical receptor and developed a novel cellular model using fluorogen-activated protein technology that allows detection of CGRP receptor internalization by flow cytometry and, for an extended time course, visualization by confocal microscopy.ResultsUsing this cell model we showed that these antagonist antibodies block both CGRP-induced cAMP signaling and CGRP receptor internalization. At least 10-fold higher concentrations of either antibody are necessary to block CGRP receptor internalization compared with cAMP accumulation in our cell model.ConclusionThese data reinforce our understanding of how monoclonal functional antagonist antibodies interfere with CGRP signaling.

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