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Magic™ Custom Ion Channel Production Service

Creative Biolabs offers one-stop, custom-oriented ion channel production services based on our leading-edge Magic™ technique platform. Currently, we have developed, optimized and commercialized multiple strategies for ion channel preparation for drug discovery, assay development, structural analysis, etc.

Ion channels are pore-forming integral membrane proteins that constitute the passage of ions across plasma membranes. In human genome there are almost 400 different ion channels locating on a broad range of cell types, governing various biophysical activities. According to ion conductivity, they can be generally classified into sodium, potassium, calcium, chloride and non-selective cation channels. In contrast to membrane pump or transporters, ion channels are only capable of passive transport down their electrochemical gradient. The opening and closing (“gating system”) are majorly governed by two methods: voltage-gated and ligand-gated. Voltage-gated ion channels are responsive to the voltage gradient across the membrane, e.g. Na+ channels, while ligand-gated ion channels are linked with functionally related receptor, and act upon ligand binding, such as ATP-gated P2X receptors.

Activation of ion channels leads to ion flux down an electrochemical concentration gradient and concomitant changes in transmembrane potential.
Figure 1. Activation of ion channels leads to ion flux down an electrochemical concentration gradient and concomitant changes in transmembrane potential. (Terstappen GC, Roncarati R, et al. 2010)

Ion channels play a critical role in regulating the electrical properties of excitable cells (e.g. neurons and myocytes), hormonal secretion, blood pressure modulation. Their dysfunction is associated with diverse pathophysiology, including neuromuscular disorders, kidney diseases, pain, hypertension, etc. During recent years, ion channels have been recognized as an important class of therapeutic target comparable to GPCRs. However, discovery and development of new ion channel drug candidates remains an arduous task due to the complex and potentially species-specific physiology, difficulties in target identification, lack of strategies to discovery and validate functional leads, and so on. The first and most crucial step is to obtain native ion channel proteins. With years of expertise in membrane protein studies and multiple state-of-the-art Magic™ membrane protein platform, Creative Biolabs now provides professional service of ion channel production. Our versatile and flexible platform offers following options:

Cell-based ion channel expression

Creative Biolabs has rich experience in ion channel heterologous expression in yeast, insect, and mammalian cell system. Exhaustive condition screening/optimization will be performed to improve the yield. With the help of powerful detergent screening platform & lipid screening platform, we can reconstitute the resulting proteins in detergent micelles, liposomes, nanodisc and polymers without compromising native structure and functionalities.

Magic™ Custom Ion Channel Production Service Cell-free ion channel expression

Cell-free machinery relies on in vitro transcription/translation to generate difficult-to-express proteins, especially like multipass ion channels. The membrane-free, detergent-free environment can greatly facilitate the chances to get functional products, which can be directly incorporated into liposome or nanodiscs. We provide prokaryotic (E. coli) and eukaryotic (wheat germ, rabbit reticulocyte lysate, insect cell, CHO/HEK) cell-free systems.

Virus-like particles and lipoparticles

Virus-like particles (VLP) represent a powerful tool for heterologous membrane protein display. Moreover, VLP-derived lipoparticle technology can capture integral ion channels in their native conformation directly from the cell surface.

Peptide derived from extracellular and intracellular domains

This is also one widely-used method for multipass proteins like ion channels. Certain peptides derived from extracellular/intracellular loops of interest can be employed as surrogate of full-length membrane proteins in immunization/antibody selection/binding assay, etc.

Ion channel stable cell line and products

Creative Biolabs has established more than 100 ion channel overexpressing stable cell lines (CHO/HEK background), which covers most of the currently hot targets, like Nav1.7. We can also construct a new stable strain upon custom request, with high expression levels up to 5 million per cell. These well-characterized, ready-to-use cell lines make perfect tools for ion channel extraction, whole-cell immunization, various cell-based assays.

In particular, we also provide purified ion channel products (reconstituted in liposomes/nanodiscs) as an alternative.

Creative Biolabs is always striving for excellence in membrane protein related studies and determined to deliver the most satisfactory service/products to our worldwide clients. Ion channel production service is one integral step in our anti-ion channel antibody development/antibody functional assay service package. In addition to ion channels, we are also professional in tailoring impeccable service of GPCRs and transporters. Please contact us for more information.

Reference

  1. Terstappen GC, Roncarati R, et al. (2010) Screening technologies for ion channel drug discovery. Future Med Chem. 2(5): 715-30.



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