title: Evaluation of recombinant monoclonal antibody SVmab1 binding to NaV1.7 target sequences and block of human NaV1.7 currents.
abstract: Identification of small and large molecule pain therapeutics that target the genetically validated voltage-gated sodium channel NaV1.7 is a challenging endeavor under vigorous pursuit. The monoclonal antibody SVmab1 was recently published to bind the NaV1.7 DII voltage sensor domain and block human NaV1.7 sodium currents in heterologous cells. We produced purified SVmab1 protein based on publically available sequence information, and evaluated its activity in a battery of binding and functional assays. Herein, we report that our recombinant SVmAb1 does not bind peptide immunogen or purified NaV1.7 DII voltage sensor domain via ELISA, and does not bind NaV1.7 in live HEK293, U-2 OS, and CHO-K1 cells via FACS. Whole cell manual patch clamp electrophysiology protocols interrogating diverse NaV1.7 gating states in HEK293 cells, revealed that recombinant SVmab1 does not block NaV1.7 currents to an extent greater than observed with an isotype matched control antibody. Collectively, our results show that recombinant SVmab1 monoclonal antibody does not bind NaV1.7 target sequences or specifically inhibit NaV1.7 current.
authors.affiliation: Neuroscience, Amgen Inc., Thousand Oaks, USA.
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.affiliation: Amgen British Columbia, Burnaby, Canada.
authors.forename: Mandy
authors.initials: M
authors.lastname: Tseng
authors.affiliation: Molecular Engineering, Amgen Inc., Cambridge, USA.
authors.forename: Linda F
authors.initials: LF
authors.lastname: Epstein
authors.affiliation: Amgen British Columbia, Burnaby, Canada.
authors.forename: Lydia
authors.initials: L
authors.lastname: Green
authors.affiliation: Amgen British Columbia, Burnaby, Canada.
authors.forename: Brian
authors.initials: B
authors.lastname: Chan
authors.affiliation: Discovery Attribute Sciences, Amgen Inc., Thousand Oaks, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Soriano
authors.affiliation: Amgen British Columbia, Burnaby, Canada.
authors.forename: Desiree
authors.initials: D
authors.lastname: Lim
authors.affiliation: Amgen British Columbia, Burnaby, Canada.
authors.forename: Oscar
authors.initials: O
authors.lastname: Pan
authors.affiliation: Amgen British Columbia, Burnaby, Canada.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Murawsky
authors.affiliation: Amgen British Columbia, Burnaby, Canada.
authors.forename: Chadwick T
authors.initials: CT
authors.lastname: King
authors.affiliation: Neuroscience, Amgen Inc., Thousand Oaks, USA.
authors.forename: Bryan D
authors.initials: BD
authors.lastname: Moyer
keywords: NaV1.7; SVmab1; antibody; electrophysiology; ion channel