Recombinant Protein Customized Service

Cloud-Clone Corp.

It is well known that after knowing the gene sequence of the protein, the gene can be cloned into the plasmid. The plasmid can be transfected into E. coli, then the E.coli is the factory for protein expression. After separation and purification, the recombinant protein can be gotten. E. coli expression system is on of the expressions for protein expression. Eukaryotic expressin systems also can be used in protein expression. Figure 1 showed the expression of recombinant protein.

Cloud-Clone Crop. provides one-stop services from genes to proteins. Customers also can choose one or two kinds of services according to their specific requirements. There are four protein expressions platform including prokaryotic expression systems (E.coli) and eukaryotic expression systems (yeast, baculovirus-insect cells, mammalian cells). We are always striving to provide the most comprehensive and high-quality service for every customer based on the continuous innovation and improvement.

Now, we will have an introduction for each expression system:

Prokaryotic expression system: prokaryotic expression system is not only the most commonly used expression system, but also the most economical protein expression system. The prokaryotic expression system is represented by the Escherichia coli expression system, which has the advantages of clear genetic background, low cost, high expression and the expressed product was easily to separate and purify. And its main disadvantage is the lack of processing mechanism of protein translation, such as the formation of disulfide bonds, protein glycosylation and the correct folding, the probability of obtaining active protein is small.

Yeast expression system: Pichia yeast is the most commonly used yeast, yeast expression system has the advantages of high expression, inducibility, glycosylation mechanism close to higher eukaryotes, easy purification of secreted protein, high density fermentation and so on. The disadvantage is that some protein products are easy to degrade and the expression is not controllable.

Baculovirus Expression Vector System: The expression host of baculovirus expression system is derived from the lawn of spodoptera frugiperda Sf9, Sf21 and trichoplusia ni High-Five, Tn-368. These cells grew rapidly, could reach a high cell density, adapted to suspension culture, and could be used as an ideal host for recombinant protein expression.

Mammalian cell expression system: compared with other systems, mammalian cell expression system has the advantage of being able to get the protein with the correct folding, and having complex N- glycosylation accurate O- glycosylation and other post-translational processing functions, therefore, the expressied products are closest to the natural proteins in molecular structure, physical and chemical properties and biological functions.

The main advantage of mammalian cell expression system and BEVS is  the protein posttranslational processing mechanism  is closest to the mechanism in vivo, the expressed protein can retain biological activity. The disadvantage is that the protein expression is usually low, the establishment of stable cell lines is difficult, and the production cost is high.

In addition, we can also provide the following value-added services for your customized protein:

1) The choice of different purification methods

2) The technology of rapid removal of endotoxin

3) The technology of tag removal

4) The technology of efficient one-step purification

5) Refolding of inclusion body

6) The technology of EASYFOLD

Conventional processes of recombinant proteins: 

Gene clone or gene synthesis→ Construction of subcloning plasmid→ Construction of expression plasmid→ Optimization of expression conditions→ Protein expression→ Protein purification→ Post processing: endotoxins removal, fusion tag removal, activity tests etc.

After many years of technology accumulation, Cloud-Clone Corp can provide more than 11000 kinds of proteins to you. If you need to customize recombinant protein, welcome to inquire us!

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