Discovery - Biology
At Icagen, we are leveraging our expertise and unique combination of technologies to develop a drug discovery platform targeting transporters. Icagen provides unique solutions for investigating and assaying transporters including our patented XRpro® technology. Our capabilities and expertise extend from cell line generation and assay development through screening and data interpretation, allowing us to address the needs of our customers, even for previously challenging systems.
Zinc (Zn2+) is an essential biological trace metal that plays structural and catalytic roles in a wide range of metalloproteins. Intracellular Zn2+ concentrations are highly regulated by a system of SLC transporters for import, export and transport into organelles. Recently, the role of Zn2+ in diabetes has been highlighted by the demonstration that mutations in SLC30A8 can reduce the risk of type II diabetes by 65%.
XRpro® technology enables the direct measurement of intracellular Zn2+, and Zn2+ transporter activity, without the need for indicator fluorophores, and with or without overexpression of target transporters.
A. XRpro® Zn Response Cultured cells (HEK-293) are titrated to show the ability of XRpro® to directly quantify intracellular Zn2+ for the number of cells grown in 96- and 384-well plate. B. Zn2+ transporter activity measured without overexpression as a function of Zn2+ concentration. Stopped-kinetic measurements measured Zn2+ uptake in HEK-293 cells expressing Zn2+ transporters at endogenous levels. Uptake rates determined from multiple time points are plotted as a function of Zn2+ concentration to determine apparent Vmax and KM values for the ensemble of endogenous transporters.
Numerous nonelectrogenic SLC symporters and antiporters operate without generating an electrical current. At Icagen, radiolabel and XRpro® flux experiments address nonelectrogenic systems through direct measurement of solute transport.
SLC12 transporters are nonelectrogenic systems responsible for maintaining intracellular chloride concentrations by coupling the transport of Cl- with Na+ and/or K+, and have been lined to multiple pathologies. Both XRpro® and 86Rb+ flux directly measure the activity of these and other nonelectrogenic systems, enabling high-throughput compound testing.
A. Direct Measurement of Ion Flux. Side-by-side determination of compound EC50 using XRpro® and 86Rb+ flux analyses. B. [Cl-] Dependence of an SLC12 family transporter XRpro® Rb+ flux analysis showing Cl- dependence of Rb+ flux for an SLC12 family member.