TY - JOUR
T1 - Dromedary immune response and specific Kv2.1 antibody generation using a specific immunization approach
AU - Hassiki, Rym
AU - Labro, Alain J
AU - Benlasfar, Zakaria
AU - Vincke, Cecile
AU - Somia, Mahmoud
AU - El Ayeb, Mohamed
AU - Muyldermans, Serge
AU - Snyders, Dirk J
AU - Bouhaouala-Zahar, Balkiss
PY - 2016/12
Y1 - 2016/12
N2 - Voltage-gated potassium (Kv) channels form cells repolarizing power and are commonly expressed inexcitable cells. In non-excitable cells, Kv channels such as Kv2.1 are involved in cell differentiation andgrowth. Due to the involvement of Kv2.1 in several physiological processes, these channels are promisingtherapeutic targets. To develop Kv2.1 specific antibody-based channel modulators, we applied a novelapproach and immunized a dromedary with heterologous Ltk- cells that overexpress the mouse Kv2.1channel instead of immunizing with channel protein fragments. The advantage of this approach is thatthe channel is presented in its native tetrameric configuration. Using a Cell-ELISA, we demonstrated theability 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 serumin modulating Kv2.1 currents. Cells that were incubated for 3 h with serum taken at Day 51 from the startof the immunization displayed a statistically significant 2-fold reduction in current density comparedto control conditions as well as cells incubated with serum from Day 0. Here we show that an immu-nization approach with cells overexpressing the Kv2.1 channel yields immune serum with Kv2.1 specificantibodies.
AB - Voltage-gated potassium (Kv) channels form cells repolarizing power and are commonly expressed inexcitable cells. In non-excitable cells, Kv channels such as Kv2.1 are involved in cell differentiation andgrowth. Due to the involvement of Kv2.1 in several physiological processes, these channels are promisingtherapeutic targets. To develop Kv2.1 specific antibody-based channel modulators, we applied a novelapproach and immunized a dromedary with heterologous Ltk- cells that overexpress the mouse Kv2.1channel instead of immunizing with channel protein fragments. The advantage of this approach is thatthe channel is presented in its native tetrameric configuration. Using a Cell-ELISA, we demonstrated theability 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 serumin modulating Kv2.1 currents. Cells that were incubated for 3 h with serum taken at Day 51 from the startof the immunization displayed a statistically significant 2-fold reduction in current density comparedto control conditions as well as cells incubated with serum from Day 0. Here we show that an immu-nization approach with cells overexpressing the Kv2.1 channel yields immune serum with Kv2.1 specificantibodies.
U2 - 10.1016/j.ijbiomac.2016.06.031
DO - 10.1016/j.ijbiomac.2016.06.031
M3 - Article
VL - 93
SP - 167
EP - 171
JO - International Journal of Biological Macromolecules
JF - International Journal of Biological Macromolecules
SN - 0141-8130
ER -