Section: bioprocessing and biological engineering gene fragment polymerization for increased yield of recombinant HIV fusion inhibitor enfuvirtide |
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Authors: | Anil Gudipudi Chitra Bajji Ravikanth Reddy Kosana Kalpana Panati Dakshayani Lomada Venkat R R Arva Tatireddigari Venkata Ramireddy Narala |
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Institution: | 1. Virchow Research Center, Hyderabad, 500 043, Andhra Pradesh, India 2. Department of Biotechnology, Government College for Men, Kadapa, Andhra Pradesh, India 3. Department Genetics & Genomics, Yogi Vemana University, Kadapa, 516 003, Andhra Pradesh, India 4. Department of Zoology, Yogi Vemana University, Kadapa, 516 003, Andhra Pradesh, India
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Abstract: | Fuzeon (Enfuvirtide, T20) is the first fusion inhibitor approved by the FDA of the USA for the treatment of HIV/AIDS in combination with other anti-retroviral drugs. Enfuvirtide is a synthetic peptide that blocks the entry of HIV into healthy host CD4 cells, which requires very high (90 mg twice daily) therapeutic doses. To increase the yield of Enfuvirtide, a gene polymerization strategy was introduced and recombinant T20 (rT20) was expressed in Escherichia coli as a five copy repeat polypeptide with a histidine-tag. The five copy rT20 was purified by Ni-affinity chromatography and cleaved to single rT20 units by cyanogen bromide. Finally, single rT20 units were purified by reversed phase chromatography giving a yield (400 mg/l) with a purity >95 %, which exhibited specific biological activity similar to Fuzeon. |
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