Dromedary immune response and specific Kv2.1 antibody generation using a specific immunization approach.
Voltage-gated potassium (Kv) channels form cells repolarizing power and are commonly expressed in excitable cells. In non-excitable cells, Kv channels such as Kv2.1 are involved in cell differentiation and growth. Due to the involvement of Kv2.1 in several physiological processes, these channels are promising therapeutic targets. To develop Kv2.1 specific antibody-based channel modulators, we applied a novel approach and immunized a dromedary with heterologous Ltk- cells that overexpress the mouse Kv2.1 channel instead of immunizing with channel protein fragments. The advantage of this approach is that the channel is presented in its native tetrameric configuration. Using a Cell-ELISA, we demonstrated the ability of the immune serum to detect Kv2.1 channels on the surface of cells that express the channel. Then, using a Patch Clamp electrophysiology assay we explored the capability of the dromedary serum in modulating Kv2.1 currents. Cells that were incubated for 3h with serum taken at Day 51 from the start of the immunization displayed a statistically significant 2-fold reduction in current density compared to control conditions as well as cells incubated with serum from Day 0. Here we show that an immunization approach with cells overexpressing the Kv2.1 channel yields immune serum with Kv2.1 specific antibodies.
Laboratoire des Venins et de Molécules Thérapeutiques, Institut Pasteur Tunis, University Tunis El Manar, 13 Place Pasteur, BP-74, 1002 Tunis, Tunisia.
Rym
R
Hassiki
Laboratory for Molecular Biophysics, Physiology, and Pharmacology, Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium.
Alain J
AJ
Labro
Service des unités animalières, Institut Pasteur Tunis, Tunisia.
Zakaria
Z
Benlasfar
Laboratory of Cellular and MolecularImmunology, VrijeUniversiteit Brussel, Brussels, Belgium.
Cécile
C
Vincke
Laboratoire des Venins et de Molécules Thérapeutiques, Institut Pasteur Tunis, University Tunis El Manar, 13 Place Pasteur, BP-74, 1002 Tunis, Tunisia.
Mahmoud
M
Somia
Laboratoire des Venins et de Molécules Thérapeutiques, Institut Pasteur Tunis, University Tunis El Manar, 13 Place Pasteur, BP-74, 1002 Tunis, Tunisia.
Mohamed
M
El Ayeb
Laboratory of Cellular and MolecularImmunology, VrijeUniversiteit Brussel, Brussels, Belgium.
Serge
S
Muyldermans
Laboratory for Molecular Biophysics, Physiology, and Pharmacology, Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium.
Dirk J
DJ
Snyders
Laboratoire des Venins et de Molécules Thérapeutiques, Institut Pasteur Tunis, University Tunis El Manar, 13 Place Pasteur, BP-74, 1002 Tunis, Tunisia; Faculté de Médecine de Tunis, Université Tunis El Manar, Tunisia. Electronic address: balkiss.bouhaouala@pasteur.rns.tn.
Balkiss
B
Bouhaouala-Zahar
D000818:Animals; D000906:Antibodies / Q000097:blood* / Q000494:pharmacology; D000917:Antibody Formation; D002162:Camelus; D002460:Cell Line; D007114:Immunization; D008297:Male; D051379:Mice; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers / Q000097:blood* / Q000494:pharmacology; D051669:Shab Potassium Channels / Q000037:antagonists & inhibitors / Q000276:immunology*
D000906:Antibodies; C496990:Kcnb1 protein, mouse; D026902:Potassium Channel Blockers; D051669:Shab Potassium Channels
Cell-ELISA; Immune response; Kv specific serum; Patch clamp