Insect cells are higher eukaryotic systems able to express large quantities of recombinant proteins with complex post-translational modifications (PTMs), such as human proteins, that cannot be produced using protein expression in bacteria. Insect cell-based protein expression constitutes a cost-effective approach for the solubilization and folding of proteins of mammalian origins with preservation of their antigenic, immunogenic and functional properties.
Together with collaborators of the Institute of Chemistry of Natural Substances (ICSN, France) and with Biospringer, Cortec Isotope has developed a straightforward, inexpensive and robust method for 15N labeled protein expression in insect cells based on labeled yeast extracts. The approach employs amino acid depleted insect cell media supplemented with 15N labeled autolysate and allows for robust 15N labeling of eukaryotic proteins. The procedure was applied to the production of a non-polymerizable mutant of actin in Spodoptera frugiperda (Sf9) cells and of fragments of eukaryotic and viral membrane fusion proteins in Drosophila Schneider 2 (S2) cells. In both cases, production yields comparable to those obtained with standard commercial media were obtained.
15N-HSQC spectrum of a 17 kDa fragment of viral protein produced in 15N labeled yeast extract (Meola et al, J. Struct. Biol. 2014)
Cortec Isotope is pleased to offer complete kits for 15N and 2H,15N labeling of protein expressed in insect cells as well as individual reagents. Each kit contains 10 g of labeled yeast extract, 1 L of amino acid depleted expression media and 270 mg of 15N Ammonium Chloride.
Cortec Isotope proposes to help you label your protein. For inquiry, please contact our labeled protein expression platform.