| Cyanovirin-N, which is an 11-KD anti-HIV protein originally isolated from extracts of a cyanobacterium, Nostoc ellipsosporum, can inhibit the Influenza A (H1N1) virus, Ebola virus. Herpes simplex virus type-1, Hepatitis C virus infection. Cyanovirin-N is a kind of the antiviral medicine due to the potent antiviral activity.Although expression systems for recombinant CVN have been established in Escherichichia coli and certain eukaryotic cells, recombinant product CV-N with this method can't meet the need of large-scale production of clinical grade CV-N for further research and development because of insufficient yield, aggregation and abnormal modification. In order to improve the production of the L-CVN, we optimized the conditions of the fermentation from somes elements including the stability of the plasmid, plasmid copy number, cultured temperature and so forth. Under these optimal conditions in a 30-L fermentor, the average expression level of soluble SUMO-CVN of three batches was up to 42.19%±3.51.We also evaluated the safety of the L-CVN and mPEG10K-ALD-LCVN by acute toxicity test in the rats and we found that the two recombitant proteins were safe under the dose of 200 mg/kg.In addition, we carried out a comparison between the immunogenicitiy of L-CVN and that of mPEG10K-ALD-LCVN to the rats. The results of these experiments showed that mPEG10K-ALD-LCVN has lower influences on the immune systems of rats than L-CVN, which indicates that mPEG10K-ALD-LCVN would be the more effective medicine with antiviral activity.Finally, we produced the anti-LCVN antibody using the purify recombinant L-CVN. IgG was purified by DEAE chromatography followed by ammonium sulfate precipitation and the purified IgG was labeled by HRP. The purified antibody with the titer reached 1:6 400 was successfully obtained with DEAE chromatograpHy followed by ammonium sulfate precipitation and the anti-CVN antibody-HRP conjugate was achieved after labeling the purified antibody with HRP.
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