A multiplatform strategy for the discovery of conventional monoclonal antibodies that inhibit the voltage-gated potassium channel Kv1.3

MAbs
Janna BednenkoPaul Colussi

Abstract

Identifying monoclonal antibodies that block human voltage-gated ion channels (VGICs) is a challenging endeavor exacerbated by difficulties in producing recombinant ion channel proteins in amounts that support drug discovery programs. We have developed a general strategy to address this challenge by combining high-level expression of recombinant VGICs in Tetrahymena thermophila with immunization of phylogenetically diverse species and unique screening tools that allow deep-mining for antibodies that could potentially bind functionally important regions of the protein. Using this approach, we targeted human Kv1.3, a voltage-gated potassium channel widely recognized as a therapeutic target for the treatment of a variety of T-cell mediated autoimmune diseases. Recombinant Kv1.3 was used to generate and recover 69 full-length anti-Kv1.3 mAbs from immunized chickens and llamas, of which 10 were able to inhibit Kv1.3 current. Select antibodies were shown to be potent (IC50<10 nM) and specific for Kv1.3 over related Kv1 family members, hERG and hNav1.5.

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Citations

Sep 1, 2018·Current Protocols in Pharmacology·Caroline S ColleyTrevor C I Wilkinson
Nov 14, 2018·Critical Reviews in Biotechnology·María E ElgueroAlejandro D Nusblat
Feb 8, 2019·Nature Reviews. Drug Discovery·Heike WulffVladimir Yarov-Yarovoy
Feb 18, 2020·Natural Product Reports·Saumya BajajK George Chandy
Jul 17, 2021·Trends in Pharmacological Sciences·Ellen GulezianSunyia Hussain
Aug 25, 2020·ACS Pharmacology & Translational Science·Seow Theng OngK George Chandy

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Methods Mentioned

BETA
confocal microscopy
Fluorescence microscopy
flow cytometry
ELISA
surface plasmon resonance
ELISAs
phage display
PCR
chip

Software Mentioned

GenScript
GraphPad Prism
Origin

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