A single-chain antibody fragment is functionally expressed in the cytoplasm of both Escherichia coli and transgenic plants

European Journal of Biochemistry

Volumn 262, Issue 2, 1999, Pages 617-624

  Tavladoraki, Paraskevi ^a,f   Girotti, Alessandra ^a   Donini, Marcello ^a   Arias, Francisco Javier ^a   Mancini, Camillo ^a,b   Morea, Veronica ^c   Chiaraluce, Roberta ^d   Consalvi, Valerio ^d   Benvenuto, Eugenio ^a,e  

^a ENEA   (Italy)

^b Dipartimento Ambiente   (Italy)

^c IRBM   (Italy)

^d SAPIENZA UNIVERSITY OF ROME   (Italy)

^e Dipartimento Innovazione   (Italy)

^f ROMA TRE UNIVERSITY   (Italy)

Author keywords

Cytosol; Disulfide bond; Intrabody; ScFv fragment

Indexed keywords

ANTIBODY;

ARTICLE; ESCHERICHIA COLI; NONHUMAN; OXIDATION REDUCTION STATE; PRIORITY JOURNAL; PROTEIN ANALYSIS; PROTEIN EXPRESSION; PROTEIN FOLDING; PROTEIN STABILITY; TRANSGENIC PLANT;

CYSTEINE; CYTOPLASM; DISULFIDES; ESCHERICHIA COLI; IMMUNOGLOBULIN VARIABLE REGION; PLANTS, GENETICALLY MODIFIED; PROTEIN DENATURATION; PROTEIN SORTING SIGNALS;

EID: 0033152614     PISSN: 00142956     EISSN: None     Source Type: Journal    
DOI: 10.1046/j.1432-1327.1999.00443.x     Document Type: Article

References (39)

1. 1. Cattaneo, A. & Biocca, S. (1997) Intracellular Antibodies: Developments and Application. Springer, New York, Berlin.
2. 2. Stieger, M., Neuhaus, G., Momma, T., Schell, J. & Kreuzaler, F. (1991) Self assembly of immunoglohulins in the cytoplasm of the alga Acetabularia mediterranea. Plant Sci. 73, 181-190.
3. 3. Carlson, J.R. (1988) A new means of inducibly inactivating a cellular protein. Mol. Cell. Biol. 8, 2638-2646.
4. 4. Cabilly, S. (1989) Growth at sub-optimal temperatures allows the production of functional, antigen-binding Fab fragments in Escherichia coli. Gene 85, 553-557.
5. 5. Biocca, S., Neuberger, M.S. & Cattaneo, A. (1990) Expression and targeting of intracellular antibodies in mammalian cells. EMBO J. 9, 101-108.
6. 6. Hiatt, A., Cafferkey, R. & Bowdish, K. (1989) Production of antibodies in transgenic plants. Nature 342, 76-78.
7. 7. Bowdish, K., Tang, Y., Hicks, J.B. & Hilvert, D. (1991) Yeast expression of a catalytic antibody with chorismate mutase activity. J. Biol. Chem. 266, 11901-11908.
8. 8. Biocca, S., Pierandrei-Amaldi, P., Campioni, N. & Cattaneo, A. (1994) Intracellular immunization with cytosolic recombinant antibodies. Bio/Technol. 12, 396-399.
9. 9. Tavladoraki, P., Benvenuto, E., Trinca, S., De Martinis, D., Cattaneo, A. & Galeffi, P. (1993) Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366, 469-472.
10. 10. Owen, M., Gandecha, A., Cockburn, B. & Whitelam, G. (1992) Synthesis of a functional anti-phytochrome single-chain Fv protein in transgenic tobacco. Bio/Technol 10, 790-794.
11. 11. Duan, L., Bagasra, O., Laughlin, M.A., Oakes, J.W. & Pomerantz, R.J. (1994) Potent inhibition of human immunodeficiency virus type 1 replication by an intracellular anti-Rev single-chain antibody. Proc. Natl Acad. Sci. USA 91, 5075-5079.
12. 12. Mhashilkar, A.M., Bagley, J., Chen, S.Y., Szilvay, A.M., Helland, D.G. & Marasco, W.A. (1995) Inhibition of HIV-1 Tat-mediated LTR transactivation and HIV-1 infection by anti-Tat single chain intrabodies. EMBO J. 14, 1542-1551.
13. 13. Fiedler, U. & Conrad, U. (1995) High-level production and long-term storage of engineered antibodies in transgenic tobacco seeds. Bio/Teehnol. 13, 1090-1093.
14. 14. Schouten, A., Roosien, I., van Engelen, F.A., de Jong, G.A.M.I., Borst-Vrenssen, A.W.M., Zilverentant, J.F., Bosch, D., Stiekema, W.J., Gommers, F.J., Schots, A. & Bakker, J. (1996) The C-terminal KDEL sequence increases the expression level of a single chain antibody designed to be targeted to both the cytosol and the secretory pathway in transgenic tobacco. Plant Mol. Biol. 30, 781-793.
15. 15. Bruyns, A.M., DeJaeger, G., DeNeve, M., De Wilde, C., Van Montagu, M. & Depicker, A. (1996) Bacterial and plant-produced scFv proteins have similar antigen-binding properties. FEBS Lett. 386, 5-10.
16. 16. Glockshuber, R., Schmidt, T. & Plückthun, A. (1992) The disulfide bonds in antibody variable domains: effects on stability, folding in vitro, and functional expression in Escherichia coli. Biochemistry 31, 1270-1279.
17. 17. Steipe, B., Schiller, B., Plückthun, A. & Steinbacher, S. (1994) Sequence statistics reliably predict stabilizing mutations in a protein domain. J. Mol. Biol. 240, 188-192.
18. 18. Knappik, A. & Plückthun, A. (1995) Engineered turns of a recombinant antibody improve its in vivo folding. Protein Engineering 8, 81-89.
19. 19. Proba, K., Honegger, A. & Plückthun, A. (1997) A natural antibody missing a cysteine in VH: consequences for thermodynamic stability and folding. J. Mol. Biol. 265, 161-172.
20. 20. Proba, K., Wörn, A. & Plückthun, A. (1998) Antibody scFv fragments without disulfide bonds made by molecular evolution. J. Mol. Biol. 275, 245-253.
21. 21. Tavladoraki, P., Franconi, R., Bradbury, A., Cattaneo, A. & Benvenuto, E. (1995) The expression of 'single chain' antibodies in transgenic plants. In Antibody Engineering (Borrebaeck, C. A. K., ed.), pp. 363-376. Oxford University Press, New York and Oxford.
22. 22. Amit, A.G., Mariuzza, R.A., Phillips, S.E.V. & Poljak, R.J. (1986) Three-dimensional structure of an antigen-antibody complex at 2.8Å resolution. Science 233, 747-753.
23. 23. Derman, A.I., Prinz, W.A., Belin, D. & Beckwith, J. (1993) Mutations that allow disulfide bond formation in the cytoplasm of Escherichia coli. Science 262, 1744-1747.
24. 24. Evan, G.I., Lewis, G.K., Ramsay, G. & Bishop, J.M. (1985) Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol. Cell. Biol. 5, 3610-3616.
25. 25. Zhu, X.-Y. & Negrutiu, I. (1991) Isolation and culture of protoplasts. In A Laboratory Guide for Cellular and Molecular Plant Biology (Negrutiu, I. & Gharti-Chhertri, G., eds), pp. 18-27. Birkhäuser, Basel, Boston and Berlin.
26. 26. Eftink, M.R. & Ghiron, C.A. (1981) Fluorescence quenching studies with proteins. Anal. Biochem. 114, 199-227.
27. 27. Lehrer, S.S. (1971) Solute perturbation of protein fluorescence. The quenching of the tryptophylfluorescence of model compounds and of lysozyme by iodide ion. Biochemistry 10, 3254-3263.
28. 28. Royer, C.A., Mann, C.J. & Matthews, C.R. (1993) Resolution of the fluorescence equilibrium unfolding profile of trpaporepressor using single tryptophan mutants. Protein Sci. 2, 1844-1852.
29. 29. Santoro, M.M. & Bolen, D.W. (1988) Unfolding free energy changes determined by the linear extrapolation method. I. Unfolding of phenylmethanesulfonyl alpha-chymotrypsin using different denaturants. Biochemistry 27, 8063-8068.
30. 30. Biocca, S., Ruberti, F., Tafani, M., Pierandrei-Amaldi, P. & Cattaneo, A. (1995) Redox state of single chain Fv fragments targeted to the endoplasmic reticulum, cytosol and mitochondria. Bio/Technol. 13, 1110-1115.
31. 31. Muesch, A., Hartmann, E., Rohde, K., Rubartelli, A., Sitia, R. & Rapoport, T.A. (1990) A novel pathway for secretory proteins? Trends Biochem. Sci. 15, 86-88.
32. 32. Lesk, A.M. & Chothia, C. (1982) Evolution of proteins formed by beta-sheets. II. The core of the immunoglobulin domains. J. Mol. Biol. 160, 325-342.
33. 33. Firek, S., Draper, J., Owen, M.R.L., Gandecha, A., Cockburn, B. & Whitelam, G.C. (1993). Secretion of a functional single-chain Fv protein in transgenic tobacco plants and cell suspension cultures. Plain Mol. Biol. 23, 861-870.
34. 34. Marasco, W.A., Haseltine, W.A. & Chen, S.Y. (1993) Design, intracellular expression, and activity of a human anti-human immunodeficiency virus type I gp120 single-chain antibody. Proc. Natl Acad. Sci. USA 90, 7889-7893.
35. 35. Pantoliano, M.W., Bird, R.E., Johnson, S., Asel, E.D., Dodd, S.W., Wood, J.F. & e Hardman, K.D. (1991). Conformational stability and ligand-binding affinity of single-chain Fv immunoglobulin fragments expressed in Escherichia coli. Biochemistry 30, 10117-10125.
36. 36. Bedzyk, W.D., Weidner, K.M., Denzin, L.K., Johnson, L.S., Hardman, K.D., Pantoliano, M.W., Asel, E.D. & Voss, E.W. Jr (1990) Immunological and structural characterization of a high affinity anti-fluorescein single-chain antibody. J. Biol. Chem. 265, 18615-18620.
37. 37. Wörn, A. & Plückthun, A. (1998) An intrinsically stable antibody scFv fragment can tolerate the loss of both disulfide bonds and fold correctly. FEBS Lett. 427, 357-361.
38. 38. Schouten, A., Roosien, J., de Boer, J.M., Wilmink, A., Rosso, M.N., Bosch, D., Stiekema, W.J., Gommers, F.J., Bakker, J. & Schots, A. (1997) Improving scFv antibody expression levels in the plant cytosol. FEBS Lett. 415, 235-241.
39. 39. Martineau, P., Jones, P. & Winter, G. (1998) Expression of an antibody fragment at high levels in the bacterial cytoplasm. J. Mol. Biol. 280, 117-127.