Rhizosecretion of a monoclonal antibody protein complex from transgenic tobacco roots

被引:48
作者
Drake, PMW [1 ]
Chargelegue, DM [1 ]
Vine, ND [1 ]
van Dolleweerd, CJ [1 ]
Obregon, P [1 ]
Ma, JKC [1 ]
机构
[1] Guys Hosp, Dept Oral Med & Pathol, Immunol Unit, GKT Dent Inst, London SE1 9RT, England
关键词
recombinant antibody; rhizosecretion; transgenic plants;
D O I
10.1023/A:1023909331482
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The secretion of a functional, full-length monoclonal antibody complex from transgenic Nicotiana tabacum roots has been demonstrated. Initially, seeds were germinated on nitrocellulose membranes and antibody secretion detected from the developing roots. Plants were then established in hydroponic culture and secretion into the growth medium measured over 25 days. Western blotting indicated that full-length antibody was present in the medium along with other fragments. Secreted antibody was shown to be functional by binding to antigen in ELISA studies. In contrast, no antibody could be detected from transgenic Nicotiana in which the same antibody was expressed as a membrane protein in the plasmalemma. These results indicate that antibody accumulation in the growth medium is genuinely caused by rhizosecretion and not cell damage. Addition of gelatin to plant growth medium markedly increased levels of antibody accumulation. The mean antibody yield per plant was calculated to be 11.7 mug per gram root dry weight per day. Rhizosecretion may be a viable alternative to agricultural production or cell culture for the generation of monoclonal antibodies in transgenic plants. It may also give rise to novel applications for antibodies expressed in plants such as removal or neutralisation of environmental pollutants and attenuation of pathogens which infect the plant via the rhizosphere.
引用
收藏
页码:233 / 241
页数:9
相关论文
共 27 条
[1]   PHYTOANTIBODIES - A GENERAL VECTOR FOR THE EXPRESSION OF IMMUNOGLOBULIN DOMAINS IN TRANSGENIC PLANTS [J].
BENVENUTO, E ;
ORDAS, RJ ;
TAVAZZA, R ;
ANCORA, G ;
BIOCCA, S ;
CATTANEO, A ;
GALEFFI, P .
PLANT MOLECULAR BIOLOGY, 1991, 17 (04) :865-874
[2]   Production of recombinant proteins in plant root exudates [J].
Borisjuk, NV ;
Borisjuk, LG ;
Logendra, S ;
Petersen, F ;
Gleba, Y ;
Raskin, I .
NATURE BIOTECHNOLOGY, 1999, 17 (05) :466-469
[3]   ANTIBODY - THE FLEXIBLE ADAPTER MOLECULE [J].
BURTON, DR .
TRENDS IN BIOCHEMICAL SCIENCES, 1990, 15 (02) :64-69
[4]   DETERMINATION OF THE PORE-SIZE OF CELL-WALLS OF LIVING PLANT-CELLS [J].
CARPITA, N ;
SABULARSE, D ;
MONTEZINOS, D ;
DELMER, DP .
SCIENCE, 1979, 205 (4411) :1144-1147
[5]   Compartment-specific accumulation of recombinant immunoglobulins in plant cells: an essential tool for antibody production and immunomodulation of physiological functions and pathogen activity [J].
Conrad, U ;
Fiedler, U .
PLANT MOLECULAR BIOLOGY, 1998, 38 (1-2) :101-109
[6]   ASSEMBLY OF AN ANTIBODY AND ITS DERIVED ANTIBODY FRAGMENT IN NICOTIANA AND ARABIDOPSIS [J].
DENEVE, M ;
DELOOSE, M ;
JACOBS, A ;
VANHOUDT, H ;
KALUZA, B ;
WEIDLE, U ;
VANMONTAGU, M ;
DEPICKER, A .
TRANSGENIC RESEARCH, 1993, 2 (04) :227-237
[7]   Intact antigen-binding MAK33 antibody and F-ab fragment accumulate in intercellular spaces of Arabidopsis thaliana [J].
DeWilde, C ;
DeNeve, M ;
DeRycke, R ;
Bruyns, AM ;
DeJaeger, G ;
VanMontagu, M ;
Depicker, A ;
Engler, G .
PLANT SCIENCE, 1996, 114 (02) :233-241
[8]   Optimization of scFv antibody production in transgenic plants [J].
Fiedler, U ;
Phillips, J ;
Artsaenko, O ;
Conrad, U .
IMMUNOTECHNOLOGY, 1997, 3 (03) :205-216
[9]   SECRETION OF A FUNCTIONAL SINGLE-CHAIN FV PROTEIN IN TRANSGENIC TOBACCO PLANTS AND CELL-SUSPENSION CULTURES [J].
FIREK, S ;
DRAPER, J ;
OWEN, MRL ;
GANDECHA, A ;
COCKBURN, B ;
WHITELAM, GC .
PLANT MOLECULAR BIOLOGY, 1993, 23 (04) :861-870
[10]   Molecular farming of pharmaceutical proteins [J].
Fischer, R ;
Emans, N .
TRANSGENIC RESEARCH, 2000, 9 (4-5) :279-299