Light-Induced Production of An Antibody Fragment and Malaria Vaccine Antigen from Chlamydomonas reinhardtii

被引:11
|
作者
Munjal, Neera [1 ]
Juliana Garzon-Sanabria, Andrea [1 ]
Quinones, Katelyn Wilson [1 ]
Gregory, James [2 ]
Nikolov, Zivko L. [1 ]
机构
[1] Texas A&M Univ, Biol & Agr Engn Dept, 2117 TAMU,Coll Stn, College Stn, TX 77843 USA
[2] Ichan Sch Med, Jaffe Food Allergy Inst, Pediat Allergy & Immunol, New York, NY 10029 USA
来源
PROCESSES | 2014年 / 2卷 / 03期
基金
美国国家科学基金会;
关键词
single-chain antibody fragment; vaccine antigen; Pfs25; Chlamydomonas reinhardtii; cultivation; recombinant protein accumulation; light induction;
D O I
10.3390/pr2030625
中图分类号
TQ [化学工业];
学科分类号
0817 ;
摘要
The eukaryotic green alga, Chlamydomonas reinhardtii, is a unique expression platform that can efficiently express complex therapeutic proteins. However, demonstrating that therapeutic molecules can be produced in quantifiable levels is essential to establish the potential of the C. reinhardtii expression system. Thus, the objective of this investigation was to determine the process conditions that could maximize C. reinhardtii biomass accumulation and induced-production of the two recombinant proteins, a single chain fragment antibody molecule (alpha CD22 scFv) and malaria vaccine antigen (Pfs25), produced in the chloroplast of C. reinhardtii. To achieve a higher production of recombinant proteins, cultivation variables of C. reinhardtii, such as mixing, light-induction time and intensity, nutrient depletion and culture age, were investigated and optimized. The optimal light-induction time was 24 h at a light intensity of 300 mu mol m(-2) s(-1). Replacement of the culture media in the late exponential growth with fresh media was beneficial to the accumulation of recombinant proteins. Optimization led to increases in the accumulation of recombinant proteins by six-fold and the recombinant protein fraction in the extracted soluble protein by two-fold.
引用
收藏
页码:625 / 638
页数:14
相关论文
共 50 条
  • [1] Modeling light-induced currents in the eye of Chlamydomonas reinhardtii
    Gradmann, D
    Ehlenbeck, S
    Hegemann, P
    JOURNAL OF MEMBRANE BIOLOGY, 2002, 189 (02): : 93 - 104
  • [2] Modeling Light-induced Currents in the Eye of Chlamydomonas reinhardtii
    D. Gradmann
    S. Ehlenbeck
    P. Hegemann
    The Journal of Membrane Biology, 2002, 189 : 93 - 104
  • [3] Identification and regulation of high light-induced genes in Chlamydomonas reinhardtii
    Im, CS
    Grossman, AR
    PLANT JOURNAL, 2002, 30 (03): : 301 - 313
  • [4] Production and recovery of multimeric malaria antigen (Pfs 25) complexes from Chlamydomonas reinhardtii chloroplast
    Wilson, Katelyn
    Munjal, Neera
    Gregory, James A.
    Mayfield, Stephen P.
    Nikolov, Zivko L.
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2014, 247
  • [5] LIGHT-INDUCED CARBONIC-ANHYDRASE EXPRESSION IN CHLAMYDOMONAS-REINHARDTII
    DIONISIOSESE, ML
    FUKUZAWA, H
    MIYACHI, S
    PLANT PHYSIOLOGY, 1990, 94 (03) : 1103 - 1110
  • [6] LIGHT-INDUCED STOP RESPONSE IN CHLAMYDOMONAS-REINHARDTII - OCCURRENCE AND ADAPTATION PHENOMENA
    HEGEMANN, P
    BRUCK, B
    CELL MOTILITY AND THE CYTOSKELETON, 1989, 14 (04): : 501 - 515
  • [7] High light-induced hydrogen peroxide production in Chlamydomonas reinhardtii is increased by high CO2 availability
    Roach, Thomas
    Na, Chae Sun
    Krieger-Liszkay, Anja
    PLANT JOURNAL, 2015, 81 (05): : 759 - 766
  • [8] Evidence for a light-induced H+ conductance in the eye of the green alga Chlamydomonas reinhardtii
    Ehlenbeck, S
    Gradmann, D
    Braun, FJ
    Hegemann, P
    BIOPHYSICAL JOURNAL, 2002, 82 (02) : 740 - 751
  • [9] UV light-induced DNA damage detection in the unicellular green alga Chlamydomonas reinhardtii
    Hercegova, Alena
    Sevcovicova, Andrea
    Galova, Eliska
    BIOLOGIA, 2008, 63 (06) : 958 - 961
  • [10] UV light-induced DNA damage detection in the unicellular green alga Chlamydomonas reinhardtii
    Alena Hercegová
    Andrea Ševčovičová
    Eliška Gálová
    Biologia, 2008, 63 : 958 - 961