Evaluation of recombinant monoclonal antibody SVmab1 binding to Na _V1.7 target sequences and block of human Na _V1.7 currents

Abstract

Identification of small and large molecule pain therapeutics that target the genetically validated voltage-gated sodium channel Na _V1.7 is a challenging endeavor under vigorous pursuit. The monoclonal antibody SVmab1 was recently published to bind the Na _V1.7 DII voltage sensor domain and block human Na _V1.7 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 Na _V1.7 DII voltage sensor domain via ELISA, and does not bind Na _V1.7 in live HEK293, U-2 OS, and CHO-K1 cells via FACS. Whole cell manual patch clamp electrophysiology protocols interrogating diverse Na _V1.7 gating states in HEK293 cells, revealed that recombinant SVmab1 does not block Na _V1.7 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 Na _V1.7 target sequences or specifically inhibit Na _V1.7 current.