Опубликовать статью

Информационное партнерство

Инфоресурсы

Контакты

«

Vol. 15, Art. 62 (pp. 779-788)    |    2014       

»

Optimization of enzymatic cleavage of peptide and protein drugs using specific proteases

Berizovskaya E.I.1, Ihalaynen A.A.1, Antohin A.M.1, Taranchenko V.F.1, Goncharov V.M.1, Mitrofanov D.A.1, Aksenov A.V.1, Rodin I.A.2, Shpigun O.A. 2

1 Federal State Unitary Enterprise "Scientific Center "The Signal", 107014, Moscow, Bolshaya Olen’a, 8. Tel. (495) 964-97-01, E-mail: eiberizovskaya@rambler.ru.
2 Federal State Educational Institution of higher professional education "Moscow State University with them. M.V. Lomonosov", Department of Chemistry, 119991, Moscow, GSP-1, Leninskie gori, 1, s.3. Tel. (495) 939-16-71.

Brief summary

Results of peptide and protein drugs enzymatic degradation study with different specific proteases – trypsin, Glu-C, Asp-N, Lys-C, Arg-C are reported. Recombinant human insulin and human chorionic gonadotropin had taken as representative example. Influence of pH, enzyme/substrate ratio, incubation temperature and incubation time was studied, optimal conditions for proteolysis of acting components was obtained.


Key words

peptides, recombinant human insulin, human chorionic gonadotropin, enzymatic digestion, specific proteases, high performance liquid chromatography.

Reference list

1. Kovaleva S.V., Isaeva I.V., Lytceva A.I. Problemi standartizacii gormonalnih preparatov peptidno-belkovoi prirodi // Ros. him. j. 2005. T. XLIX, № 1. S. 135-145.


2. British Pharmacopeia 2014, London: Bernan, 2013. 5860 r.


3. Evropeiskaya farmakopeya 7-oe izdanie. T.1. M.: Remediym, 2011, 1812 s.


4. Evropeiskaya farmakopeya 7-oe izdanie. T.2. Ch. 1. M.: Remediym, 2011, 3176 s.


5. Evropeiskaya farmakopeya 7-oe izdanie. T.2. Ch. 2. M.: Remediym, 2011, 4504 s.


6. United States Pharmacopeia: USP 27-NF 22. Rockville: Inc., 2009. 3013 p.


7. Japanese Pharmacopoeia, 16th edition. [Elektronnii resyrs] // sait. URL: http://jpdb.nihs.go.jp/jp16e/jp16e.pdf (data obrasheniya: 29.09.2014 g.).


8. Fenn J., Mann M., Meng C. Electrospray Ionization for Mass Spectrometry of Large Biomolecules // Science. 1989. Vol. 64, № 4926. P. 246.


9. Fenn J., Mann M., Meng C. Electrospray Ionization-Principles and Practice // Mass Spectrom Rev. 1990. Vol. 9, № 1. P. 37-70.


10. Wong S., Meng C., Fenn J. Multiple Charging in Electrospray Ionization of Poly(Ethylene Glycols) // J. Phys. Chem. 1988. Vol. 92, № 2. P. 546-550.


11. Wilm M., Mann M. Analytical properties of the nanoelectrospray ion source // Anal. Chem. 1996. Vol. 68, № 1. P. 1-8.


12. Tanaka K., Waki H., Ido Y. Protein and Polymer Analyses up to m/z 100 000 by Laser Ionization Time-of flight Mass Spectrometry // Rapid Commun Mass Spectrom. 1988. Vol. 2, № 20. P. 151-153.


13. Karas M, Hillenkamp F. Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons // Anal Chem. 1988. Vol. 60, № 20. P. 2299-2301.


14. Hillenkamp F., Karas M. Matrix-assisted laser desoption/ionization, an experience // Int. J. Mass Spectrom. 2000. Vol. 200, № 1-3. P. 71-77.


15. Bogdanov V., Smith R. Proteomics by FTICR Mass-Spectrometry: Top-Down and Bottom-Up // Mass Spectrom Rev. 2005. Vol. 24, № 2. P. 168- 200.


16. Aebersold R, Mann M. Mass spectrometry-based proteomics // Nature. 2003. Vol. 422, № 6928. P. 198-207.


17. Washburn M., Wolters D., Yates J. 3rd. Large-scale analysis of the yeast proteome by multidimensional protein identification technology // Nat. Biotechnol. 2001. Vol. 19, № 3. P. 242-247.