Recombinant Human Butyrylcholinesterase As a New-Age Bioscavenger Drug: Development of the Expression System

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Butyrylcholinesterase (BChE) is a serine hydrolase (EC 3.1.1.8) which can be found in most animal tissues. This enzyme has a broad spectrum of efficacy against organophosphorus compounds, which makes it a prime candidate for the role of stoichiometric bioscavenger. Development of a new-age DNA-encoded bioscavenger is a vival task. Several transgenic expression systems of human BChE were developed over the past 20 years; however, none of them has been shown to make economic sense or has been approved for administration to humans. In this study, a CHO-based expression system was redesigned, resulting in a significant increase in the production level of functional recombinant human butyrylcholinesterase as compared to the hitherto existing systems. The recombinant enzyme was characterized with Elman and ELISA methods.

About the authors

D. G. Ilyushin

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Author for correspondence.
Email: IlyushinDenis@gmail.com
Россия

O. M. Haertley

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

T. V. Bobik

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

O. G. Shamborant

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

E. A. Surina

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

V. D. Knorre

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

P. Masson

Département de Toxilogie, Centre de Recherches du Servise de Sante des Armées

Email: IlyushinDenis@gmail.com
Франция

I. V. Smirnov

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

A. G. Gabibov

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

N. A. Ponomarenko

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry

Email: IlyushinDenis@gmail.com
Россия

References

  1. Jbilo O., L’Hermite Y., Talesa V., Toutant J.P., Chatonnet A. // Eur. J. Biochem. 1994. V. 225. P. 115-124.
  2. Mesulam M. // Neuroscience. 2002. V. 110. P. 627-639.
  3. Saxena A., Sun W., Fedorko J.M., Koplovitz I., Doctor B.P. // Biochem. Pharmacol. 2011. V. 81. P. 164-169.
  4. Lockridge O., Schopfer L.M., Winger G., Woods J.H. // J. Med. Chem. Biol. Radiol. Def. 2005. V. 3. P. 1-23.
  5. Raveh L. // Toxicol. Appl. Pharmacol. 1997. V. 145. P. 43-53.
  6. Patrick M., Daniel R. // Acta Naturae. 2009. V. 1. № 1. P. 68-78.
  7. Masson P., Nachon F., Broomfield C.A., Lenz D.E., Verdier L., Schopfer L.M., Lockridge O. // Chem. Biol. Interact. 2008. V. 175. P. 273-280.
  8. Lockridge O., Bartels C.F., Vaughan T.A., Wong C.K., Norton S.E., Johnson L.L. // J. Biol. Chem. 1987. V. 262. P. 549-557.
  9. Lenz D.E., Yeung D., Smith J.R., Sweeney R.E., Lumley L.A., Cerasoli D.M. // Toxicology. 2007. V. 233. P. 31-39.
  10. Masson P., Carletti E., Nachon F. // Protein Pept. Lett. 2009. V. 16. P. 1215-1224.
  11. Geyer B.C., Kannan L., Cherni I., Woods R.R., Soreq H., Mor T.S. // Plant Biotechnol. J. 2010. V. 8. P. 873-886.
  12. Masson P., Steve A., Philippe P.-T., Oksana L. // Multidisciplinary approaches to cholinesterase functions. Expression and refoldin of functional human butyrylcholinesterase in E. coli. N.Y.: Plenum Press, 1992.
  13. Kishnani P.S., Corzo D., Nicolino M., Byrne B., Mandel H., Hwu W.L., Leslie N., Levine J., Spencer C., McDonald M., et al. // Neurology. 2007. V. 68. P. 99-109.
  14. Schwartz R.S., Abildgaard C.F., Aledort L.M., Arkin S., Bloom A.L., Brackmann H.H., Brettler D.B., Fukui H., Hilgartner M.W., Inwood M.J. // N. Engl. J. Med. 1990. V. 323. P. 1800-1805.
  15. White G.C., Beebe A., Nielsen B. // Thromb. Haemost. 1997. V. 78. P. 261-265.
  16. Jacobs L.D., Cookfair D.L., Rudick R.A., Herndon R.M., Richert J.R., Salazar A.M., Fischer J.S., Goodkin D.E., Granger C.V., Simon J.H., et al. // Ann. Neurol. 1996. V. 39. P. 285-294.
  17. Barngrover D. // J. Biotechnol. 2002. V. 95. P. 280-282.
  18. Tabuchi H., Sugiyama T., Tanaka S., Tainaka S. // Biotechnol. Bioeng. 2010. V. 107. P. 998-1003.
  19. Porter A.J., Dickson A.J., Racher A.J. // Biotechnol. Progr. 2010. V. 26. P. 1446-1454.
  20. Strnad J., Brinc M., Spudić V., Jelnikar N., Mirnik L., Carman B., Kravanja Z. // Biotechnol. Progr. 2010. V. 26. P. 653-663.
  21. Mizushima S., Nagata S. // Nucl. Acids Res. 1990. V. 18. P. 5322.
  22. Boshart M., Weber F., Jahn G., Dorsch-Häsler K., Fleckenstein B., Schaffner W. // Cell. 1985. V. 41. P. 521-530.
  23. Nachon F., Nicolet Y., Viguié N., Masson P., Fontecilla-Camps J.C., Lockridge O. // Eur. J. Biochem. 2002. V. 269. P. 630-637.
  24. Li H., Schopfer L.M., Masson P., Lockridge O. // Biochem. J. 2008. V. 411. P. 425-432.
  25. Altamirano C.V., Lockridge O. // Biochemistry. 1999. V. 38. P. 13414-13422.
  26. Huang Y.-J., Huang Y., Baldassarre H., Wang B., Lazaris A., Leduc M., Bilodeau A.S., Bellemare A., Côté M., Herskovits P., Touati M., Turcotte C., Valeanu L., et al. // Proc. Natl. Acad. Sci. USA. 2007. V. 104. P. 13603-13608.
  27. Geyer B.C., Kannan L., Garnaud P.-E., Broomfield C.A., Cadieux C.L., Cherni I., Hodgins S.M., Kasten S.A., Kelley K., Kilbourne J., Oliver Z.P., Otto T.C., Puffenberger I., et al. // Proc. Natl. Acad. Sci. USA. 2010. V. 107. P. 20251-20256.
  28. Freshney R.I. Culture of animal cells. Oxford, N.Y.: Wiley-Blackwell, 2005. P. 642.
  29. Duysen E.G., Bartels C.F., Lockridge O. // J. Pharmacol. Exp. Ther. 2002. V. 302. P. 751-758.
  30. Sambrook J., Russell D.W. Molecular Cloning. Cold Spring Harbor. N.Y.; Cold Spring Harbor Lab. Press, 2001.
  31. Sambrook J., Fritsch E.F., Maniatis T. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor. N.Y.; Cold Spring Harbor Lab. Press, 1989.
  32. Schlesinger N., Baker D.G., Schumacher H.R. // J. Rheumatol. 1997. V. 24. P. 1018-1019.
  33. Harlow E., Harlow E., Lane D. Antibodies: A Laboratory Manual. Cold Spring Harbor. N.Y.; Cold Spring Harbor Lab. Press, 1988.
  34. Ausubel F.M. // Current Protocols. 2002. Unit 2.7.
  35. Ellman G.L., Courtney K.D., Anders V., Feather-Stone R.M. // Biochem. Pharmacol. 1961. V. 7. P. 88-95.
  36. Karnovsky M.J., Roots L. // J. Histochem. Cytochem. 1964. V. 12. P. 219-221.
  37. Xie W., Altamirano C.V., Bartels C.F., Speirs R.J., Cashman J.R., Lockridge O. // Mol. Pharmacol. 1999. V. 55. P. 83-91.
  38. Bartels C.F., Jensen F.S., Lockridge O., van der Spek A.F., Rubinstein H.M., Lubrano T., La Du B.N. // Am. J. Hum. Genet. 1992. V. 50. P. 1086-1103.
  39. Wang Y., Boeck A.T., Duysen E.G., van Keuren M., Saunders T.L., Lockridge O. // Toxicol. Appl. Pharmacol. 2004. V. 196. P. 356-366.
  40. Preininger A., Schlokat U., Mohr G., Himmelspach M., Stichler V., Kyd-Rebenburg A., Plaimauer B., Turecek P.L., Schwarz H.P., Wernhart W., Fischer B.E., Dorner F. // Cytotechnology. 1999. V. 30. P. 1-15.
  41. Wajih N., Hutson S.M., Owen J., Wallin R. // J. Biol. Chem. 2005. V. 280. P. 31603-31607.
  42. Jossé L., Smales C.M., Tuite M.F. // Biotechnol. Bioeng. 2010. V. 105. P. 556-566.
  43. Meleady P., Henry M., Gammell P., Doolan P., Sinacore M., Melville M., Francullo L., Leonard M., Charlebois T., Clynes M. // Proteomics. 2008. V. 8. P. 2611-2624.
  44. Roncarati R., Seredenina T., Jow B., Jow F., Papini S., Kramer A., Bothmann H., Dunlop J., Terstappen G.S. // Assay Drug Dev. Technol. 2008. V. 6. P. 181-193.
  45. Chilukuri N., Sun W., Naik R.S., Parikh K., Tang L., Doctor B.P., Saxena A. // Chem. Biol. Interact. 2008. V. 175. P. 255-260.
  46. Jain S., Hreczuk-Hirst D.H., McCormack B., Mital M., Epenetos A., Laing P., Gregoriadis G. // Biochim. Biophys. Acta. 2003. V. 1622. P. 42-49.

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Copyright (c) 2013 Ilyushin D.G., Haertley O.M., Bobik T.V., Shamborant O.G., Surina E.A., Knorre V.D., Masson P., Smirnov I.V., Gabibov A.G., Ponomarenko N.A.

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