Interferon α2b In E.coli Of Induced The Expression Extracted And Purified

The interferon is a kind of cytokine that has multiple biologicalactivities, it significantly adjusts organism immunity function, enhancesanti-virus, anti-tumor functions and so on;interferon in clinical practice hasbeen more than 40 years. Recombinant human Interferon α2b is one kind ofanti-virus, anti-tumor and organism immunity enhancement broad-spectrummedicine. It is used in treating diseases such as hepatitis B, herpes zoster,condyloma acuminatum, leukemia, malignant tumor and so on. At present, thisproduct has been developed and put into production in many domesticfactories, but the cost is high, moreover because of complex operation andlong production cycle, it is not suitable for large-scale production. Becausethere is no glycosylation sites in IFN;it can be well represented by coli, thebiological activity between coli representation and the natural interferon arevery close, also the coli has the advantages of high output, simple technique,low cost. In order to enhance the quality and quantity of interferon α2b, andreduce the cost, we have constructed the engineered bacteria, and fermentedthe interferon α2b engineered bacteria, induced the expression, extracted andpurified interferon α2b.This research carried out two PCR that produced IFN -2b gene,increased the A+T content of 5'nd;changed 4 coli rare codons to theirpreferred codons, inserted IFN-2b gene into the end of PRPL of PBV-220,transformed coli DH5, constructed efficient engineeringstrain of the interferon α2b gene, this strain not only expresses highquantity, moreover interferon α2b exists in the cytoplasm as inclusion body,inclusion body helps to prevent the bacterium proteinase to disturb and digestinterferon α2b, benefits the separation and purification processes. Based onmany times of experiments, we have successfully established a set of feasibleand practical strains fermentation, inclusion body extraction, denaturation,renaturation and purification, it can produce good interferon α2b productwithout the need of the monoclonal antibody chromatographic, Its eachexamination result conforms to《 Pharmacopoeia of peopme republic of china》2005 edition third.