Мақаланы E-mail арқылы жіберу Synthesis in Escherichia coli and Characterization of Human Recombinant Erythropoietin with Additional Heparin-Binding Domain
- Авторлар: Karyagina A.1,2,3, Grunina T.1, Poponova M.1, Orlova P.1, Manskikh V.1,3, Demidenko A.1, Strukova N.1, Manukhina M.1, Nikitin K.1, Lyaschuk A.1, Galushkina Z.1, Cherepushkin S.4, Polyakov N.1,5, Solovyev A.1, Zhukhovitsky V.1, Tretyak D.6, Boksha I.1,7, Gromov A.1, Lunin V.1,2
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Мекемелер:
- Gamaleya National Research Center of Epidemiology and Microbiology
- All-Russia Research Institute of Agricultural Biotechnology
- Belozersky Institute of Physical and Chemical Biology
- State Research Institute of Genetics and Selection of Industrial Microorganisms
- Vernadsky Institute of Geochemistry and Analytical Chemistry
- Moscow Technological University (Lomonosov Institute of Fine Chemical Technologies)
- Research Center of Mental Health
- Шығарылым: Том 83, № 10 (2018)
- Беттер: 1207-1221
- Бөлім: Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151738
- DOI: https://doi.org/10.1134/S0006297918100061
- ID: 151738
Дәйексөз келтіру
Ашық рұқсат
Рұқсат берілді
Тек жазылушылар үшін
Аннотация
Recombinant human erythropoietin (EPO) with additional N-terminal heparin-binding protein domain (HBD) from bone morphogenetic protein 2 was synthesized in Escherichia coli cells. A procedure for HBD-EPO purification and refolding was developed for obtaining highly-purified HBD-EPO. The structure of recombinant HBD-EPO was close to that of the native EPO protein. HBD-EPO contained two disulfide bonds, as shown by MALDI-TOF mass spectrometry. The protein demonstrated in vitro biological activity in the proliferation of human erythroleukemia TF-1 cell test and in vivo activity in animal models. HBD-EPO increased the number of reticulocytes in the blood after subcutaneous injection and displayed local angiogenic activity after subcutaneous implantation of demineralized bone matrix (DBM) discs with immobilized HBD-EPO. We developed a quantitative sandwich ELISA method for measuring HBD-EPO concentration in solution using rabbit polyclonal serum and commercial monoclonal anti-EPO antibodies. Pharmacokinetic properties of HBD-EPO were typical for bacterially produced EPO. Under physiological conditions, HBD-EPO can reversibly bind to DBM, which is often used as an osteoplastic material for treatment of bone pathologies. The data on HBD-EPO binding to DBM and local angiogenic activity of this protein give hope for successful application of HBD-EPO immobilized on DBM in experiments on bone regeneration.
Негізгі сөздер
Авторлар туралы
A. Karyagina
Gamaleya National Research Center of Epidemiology and Microbiology; All-Russia Research Institute of Agricultural Biotechnology; Belozersky Institute of Physical and Chemical Biology
Хат алмасуға жауапты Автор.
Email: akaryagina@gmail.com
Ресей, Moscow, 123098; Moscow, 127550; Moscow, 119991
T. Grunina
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
M. Poponova
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
P. Orlova
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
V. Manskikh
Gamaleya National Research Center of Epidemiology and Microbiology; Belozersky Institute of Physical and Chemical Biology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098; Moscow, 119991
A. Demidenko
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
N. Strukova
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
M. Manukhina
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
K. Nikitin
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
A. Lyaschuk
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
Z. Galushkina
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
S. Cherepushkin
State Research Institute of Genetics and Selection of Industrial Microorganisms
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 117545
N. Polyakov
Gamaleya National Research Center of Epidemiology and Microbiology; Vernadsky Institute of Geochemistry and Analytical Chemistry
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098; Moscow, 119334
A. Solovyev
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
V. Zhukhovitsky
Gamaleya National Research Center of Epidemiology and Microbiology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
D. Tretyak
Moscow Technological University (Lomonosov Institute of Fine Chemical Technologies)
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 119571
I. Boksha
Gamaleya National Research Center of Epidemiology and Microbiology; Research Center of Mental Health
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098; Moscow, 115522
A. Gromov
Gamaleya National Research Center of Epidemiology and Microbiology
Хат алмасуға жауапты Автор.
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098
V. Lunin
Gamaleya National Research Center of Epidemiology and Microbiology; All-Russia Research Institute of Agricultural Biotechnology
Email: alexander.v.gromov@gmail.com
Ресей, Moscow, 123098; Moscow, 127550
