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

MAbs. 2018 May/Jun;10(4):636-650. doi: 10.1080/19420862.2018.1445451. Epub 2018 Apr 2.

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.

Keywords: Kv1.3; Tetrahymena; antibody; voltage-gated ion channel.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Camelids, New World
  • Chickens
  • Drug Discovery / methods*
  • Humans
  • Kv1.3 Potassium Channel / antagonists & inhibitors*
  • Recombinant Proteins
  • Tetrahymena thermophila

Substances

  • Antibodies, Monoclonal
  • Kv1.3 Potassium Channel
  • Recombinant Proteins

Grant support

This work was supported, in part, by the Juvenile Diabetes Research Foundation T1D Fund-PI Paul Colussi, PhD, pcolussi@tetragenetics.com.