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Mempro™ Cell-Free Lipocalin Production

Creative Biostructure can provide custom Mempro™ protein production services for lipocalins using cell-free expression system.

Mempro™ cell-free protein production system is the progressive platform to produce intergral membrane proteins which can bypass the limitations that encountered in cell-based protein production system, such as host cell toxicity, low yield and solubilization and purification using detergents.

Members of lipocalin protein family, which contain a buried structural pocket as a binding site, can form a different group of ligand-binding proteins that play important roles in transprot, storage, or sequestration of a variety of small hydrophobic ligands. Most lipocalins possess a signal peptide at N-terminus, and several disulfide bridges that contribute to constraint of the overall structure through keep sabilization of the N- and C-terminal regions of the lipocalin. It has been reported that lipocalins are associated with a variety of biological processes, such as cancer cell interactions, biological prostaglandin synthesis, immune response, pheromone transport and retinoid binding.

MemproTM Cell-Free Lipocalin Production Figure 1. The structure of human lipocalin. (Acc. Chem., 2015)

Creative Biostructure has extensive experience in high-yield lipocalin using cell-free protein expression system, we can provide various strategies for Mempro™ cell-free protein production with post-tranlational modifications, including:

Among Mempro™ cell-free protein production platform, E. coli S30 extract is popular used for lipocalin production. Wheat germ extract can provide eukaryotic translation system for the production of lipocalins. Baculovirus-based expression system derived from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for lipocalin expression. Notably, crude rabbit reticulocyte lysate is the most important tool used for eukaryotic cell-free membrane protein expression system.

With the Mempro™ cell-free protein production platform, Creative Biostructure can express, purify, solubilize and crystallize lipocalins to study interactions with other proteins for molecular recognition. Mempro™ cell-free protein production has remarkable advantages, including:

  • Very high yield;
  • Low cost;
  • Lack of toxicity;
  • Incorporation of non-natural amino acids into protein structure;
  • Relatively tolerant of additives.

Creative Biostructure can provide various custom Mempro™ membrane protein production services. Please feel free to contact us for more information.

References:
A. Schiefner and A. Skerra (2015). The Menagerie of Human Lipocalins: A Natural Protein Scaffold for Molecular Recognition of Physiological Compounds. Acc. Chem., 48(4): 976–985.
Lipocalin. (https://en.wikipedia.org/wiki/Lipocalin)
M. D. Ganfornina, et al. (1999). A phylogenetic analysis of the lipocalin protein family. Mol. Biol. Evol., 17(1): 114-126.


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