Bacteria protein expression system has been the most popular means of producing recombinant proteins as bacterium is fast growing, easy genetic manipulation, high yields, adapted to scale-up production and low overall cost. E. coli is a well-established host and widely used since ample knowledge has been acquired about its genetics, genome sequence and physiology. Therefore, E. coli expression system is an important system to produce many different types of proteins, including antibodies.
Creative Biolabs offers ready-to-use competent cells for expression of antibodies in E. coli and we have established several systems, for instance, T7 RNA polymerase system, for producing antibodies. T7 RNA polymerase gene promoter and an expression vector containing a gene of interest were constructed and introduced into a T7 expression host (E. coli). When inducer is added, T7 RNA polymerase is expressed and transcribes the gene of interest and the antibody can be obtained by a series of purification steps.
Creative Biolabs has a long history of producing a wide variety of recombinant antibodies. We have developed state-of-the-art bacterial expression platform to express customized antibodies. Even those antibodies that contain complex disulfide bonds or any other hard to dissolve components, they can be expressed at a decent level. We have established a track record for producing high-quality, correctly folded antibody fragments (including Fabs and scFv).
Another characteristic of our bacterial expression platform is the parallel expression. This high throughput expression technique highly speeds up protein production. We are able to construct a variety of vectors containing different tags and/or fusion partners to express different types of antibodies in parallel. For hard to express antibodies, we use the strategy of constructing a specific antibody protein in a standard vector and express them in a number of different E. coli host strains. This approach not only speeds up expression time but also results in a larger number of successfully expressed proteins.
As we know, prokaryotic protein expression systems have some disadvantages, such as no post-translational modifications, inclusion body possible and presence of endotoxins. In order to eliminate the guesswork of no post-translational modifications for antibody production, we optimize your expressed antibodies by refolded with our proprietary FoldEZ™ protein refolding technology. This technology evaluates target antibody's biochemical and biophysical properties and selects the optimal strategy based on every antibody's properties. 20 different refolding buffers were evaluated to determine the optimal refolding conditions. We provide refolded protein at purity of 95% or greater.
In order to eliminate the bacteria endotoxins, we use gel clot or chromogenic methods to detect the endotoxins and remove them by Polymyxin B (PMB) - affinity-based method, size exclusion chromatography and ion exchange chromatography. Our methods allow highly efficient removal of endotoxin down to 0.1 EU/mL.
Our well-developed bacterial expression system is a valuable tool for the expression of complex antibodies. For more detailed information, please feel free to contact us or directly sent us an inquiry.
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