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Magic™ Cell-free Protein Expression in Insect Cell Lysate

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Creative Biolabs is a global pioneer and leader in cell-free membrane protein expression using insect system. With years of experience and endeavor, we have built a unique Magic™ cell-free protein expression platform based on various prokaryotic and eukaryotic systems, such as E. coli, wheat germ, CHO, and HEK. Our experts have earned great reputation from our global customers by successfully accomplishing plenty of challenging projects during the past decade.

During the past decades, the cell-free expression has proven to be a promising platform allowing the synthesis of many different protein classes including membrane proteins, toxic proteins, and proteins with post-translational modifications. Many thorny problems connected with a cell-based expression of proteins can be solved by the use of tailor-made cell-free expression systems. Recently, more and more cell-free systems evolved as a promising solution for various difficult-to-express proteins, including but not restricted to:


Each of them has their own unique characteristics, and selection of the appropriate system is very critical, depending on protein type, protein function or structure, desired yield along with many other factors.

According to different cellular origin, insect cells can be classified into 3 major types including Spodoptera frugiperda cells (e.g. Sf9 and Sf21), Cabbage looper cells (e.g. Hi5), and Drosophila melanogaster cells (e.g. S2). As is known, insect cell-based system has long been the prevailing choice for expressing many eukaryotic proteins, especially integral membrane proteins like GPCRs. However, for some particular target, cellular expression can have many inevitable limitations. Hence, protein expression performed in insect cell lysates manifests as a very promising alternative. This cell-free system contains endogenous microsomal vesicles, which enables the translocation of synthesized target proteins into the lumen of the insect vesicles – especially in case of membrane proteins, their inserting into a natural membrane scaffold.

Representative pictures of Spodoptera frugiperda, Cabbage looper and Drosophila melanogaster Figure 1. Representative pictures of Spodoptera frugiperda, Cabbage looper and Drosophila melanogaster (from left to right).

Compared with E. coli, it is better compatible with eukaryotic proteins. Besides, insect cells can also endure large heterogeneous proteins with higher expression levels than conventional CHO/human cell-based systems, but fewer misfolded proteins than the prokaryotic system. Creative Biolabs has established an optimized cell-free lysate with following features:


Creative Biolabs has extensive expertise in membrane proteins expression based on our high-quality cell-free expression systems. With optimized systems and solid technological base, fast delivery and professional support are guaranteed. Our experienced experts are specialized in tailoring the most suitable and efficient plans according to different custom demands. Please feel free to inquiry us for more information.

References

  1. Ezure T, Suzuki T, Shikata M, et al. (2010) A cell-free protein synthesis system from insect cells. Cell-Free Protein Production: Methods and Protocols, 31-42.
  2. Alkhalfioui F, Logez C, Bornert O, Wagner R (2011) Production of Membrane Proteins - Strategies for Expression and Isolation (Robinson AS, ed), Wiley-VCH: 1.



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