title: Gene fragment polymerization for increased yield of recombinant HIV fusion inhibitor enfuvirtide. [corrected].
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.
authors: Virchow Research Center, Hyderabad, 500 043, Andhra Pradesh, India.
authors: Anil
authors: A
authors: Gudipudi
authors: Chitra
authors: C
authors: Bajji
authors: Ravikanth Reddy
authors: RR
authors: Kosana
authors: Kalpana
authors: K
authors: Panati
authors: Dakshayani
authors: D
authors: Lomada
authors: Venkat R R
authors: VR
authors: Arva Tatireddigari
authors: Venkata Ramireddy
authors: VR
authors: Narala
mesh_terms: D019380:Anti-HIV Agents / Q000378:metabolism* / Q000494:pharmacology; D019677:Artificial Gene Fusion; D053898:Biosynthetic Pathways / Q000235:genetics*; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D000077560:Enfuvirtide; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D006868:Hydrolysis; D060847:Metabolic Engineering; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D019380:Anti-HIV Agents; D015700:HIV Envelope Protein gp41; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D000077560:Enfuvirtide