Recombinant approach for the production of HIV fusion inhibitor Enfuvirtide using Escherichia coli

被引:4
|
作者
Kosana, Ravikanth Reddy [1 ,2 ]
Bajji, Chitra [1 ]
Kanumuri, Radha Madhavi [1 ]
Panati, Kalpana [3 ]
Mangamoori, Lakshmi Narasu [2 ]
Tummuru, Muralikrishna Reddy [1 ]
Narala, Venkata Ramireddy [4 ]
机构
[1] Virchow Res Ctr, Hyderabad 500043, AP, India
[2] Jawaharlal Nehru Technol Univ, Ctr Biotechnol, Hyderabad 500085, AP, India
[3] Govt Degree Coll Men, Dept Biotechnol, Kadapa, AP, India
[4] Yogi Vemana Univ, Dept Zool, Kadapa 516003, AP, India
关键词
Enfuvirtide; Fusion protein expression; Recombinant T20; SDS solubilization; GLUTATHIONE-S-TRANSFERASE; HUMAN GROWTH-HORMONE; PROTEIN; INFECTION; PURIFICATION; EXPRESSION; GP41;
D O I
10.1016/j.pep.2013.12.002
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The use of antiretroviral drugs is gaining importance in the recent past for the treatment of human immunodeficiency virus infection. Enfuvirtide (T20) is one of the fusion inhibitors, inhibits the fusion between the virus and healthy target CD4 cells. The treatment with T20 involves very high therapeutic dose. In addition to its high dose, production of T20 by synthetic methods is expensive and cumbersome. We report an alternative recombinant approach for the production of the T20 peptide through a novel short fusion-tag expression system. This expression system consists of the hydrophobic region of growth hormone (GH) as the fusion tag, a factor Xa cleavage site upstream to the T20. The fusion protein was expressed in Escherichia coli as inclusion bodies. We also report here, a simple and an efficient downstream strategy for the purification of recombinant T20 peptide (rT20). Our study is the first to demonstrate a novel approach using GH fusion tag, ensured the peptide stability, for the production of rT20 which yields more than 250 mg/L with 98% purity. The biological activity of the rT20 is comparable to its synthetic counterpart. Thus, this novel approach could be an alternate method of choice for production of therapeutically important small peptides. (C) 2013 Elsevier Inc. All rights reserved.
引用
收藏
页码:136 / 142
页数:7
相关论文
共 50 条
  • [1] Enfuvirtide: the first HIV fusion inhibitor
    Lazzarin, A
    EXPERT OPINION ON PHARMACOTHERAPY, 2005, 6 (03) : 453 - 464
  • [2] Enfuvirtide: A fusion inhibitor for the treatment of HIV infection
    Fung, HB
    Guo, Y
    CLINICAL THERAPEUTICS, 2004, 26 (03) : 352 - 378
  • [3] Production of recombinant human glucagon in Escherichia coli by a novel fusion protein approach
    Kim, DY
    Shin, NK
    Chang, SG
    Shin, HC
    BIOTECHNOLOGY TECHNIQUES, 1996, 10 (09) : 669 - 672
  • [4] Resistance to enfuvirtide, the first HIV fusion inhibitor
    Greenberg, ML
    Cammack, N
    JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY, 2004, 54 (02) : 333 - 340
  • [5] Enfuvirtide, an HIV-1 fusion inhibitor
    Carr, A
    NEW ENGLAND JOURNAL OF MEDICINE, 2003, 349 (18): : 1770 - 1770
  • [6] Erratum to: Gene fragment polymerization for increased yield of recombinant HIV fusion inhibitor Enfuvirtide
    Anil Gudipudi
    Chitra Bajji
    Ravikanth Reddy Kosana
    Kalpana Panati
    Dakshayani Lomada
    Venkat R. R. Arva Tatireddigari
    Venkata Ramireddy Narala
    Biotechnology Letters, 2014, 36 : 1771 - 1771
  • [7] Enfuvirtide, an HIV-1 fusion inhibitor - Reply
    Lalezari, JP
    Lazzarin, A
    Salgo, M
    NEW ENGLAND JOURNAL OF MEDICINE, 2003, 349 (18): : 1771 - 1771
  • [8] Enfuvirtide: First fusion inhibitor for treatment of HIV infection
    Jamjian, MC
    McNicholl, IR
    AMERICAN JOURNAL OF HEALTH-SYSTEM PHARMACY, 2004, 61 (12) : 1242 - 1247
  • [9] Modelling the effects of adherence to the HIV fusion inhibitor enfuvirtide
    Lou, Jie
    Smith, Robert J.
    JOURNAL OF THEORETICAL BIOLOGY, 2011, 268 (01) : 1 - 13
  • [10] Enfuvirtide, the first fusion inhibitor to treat HIV infection
    Poveda, E
    Briz, V
    Soriano, V
    AIDS REVIEWS, 2005, 7 (03) : 139 - 147