Russian Journal of Biotechnology Articles archive Archive 2015 N 1 MonoPEGylated Human Interferon beta-1a Preparation: Optimization of Conditions for N-Terminal Pegylation

MonoPEGylated Human Interferon beta-1a Preparation: Optimization of Conditions for N-Terminal Pegylation

Автор: D.V. Korzhavin, T.V. Chernovskaya, Yu.G. Efanov, E.G. Rudenko, R.A. Ivanov, A.B. Pshenichnikova, and V.I. Shvets

Страница: 49-60

 

MonoPEGylated Human Interferon beta-1a Preparation: Optimization of Conditions for N-Terminal Pegylation

Biotekhnologiya, 2015, N 1, P. 49-60

UDC 577.112.4

Section:  “Biologicals Technology”

 

D.V. Korzhavin 1, 2, *,  T.V. Chernovskaya 1,  Yu.G. Efanov 1,  E.G. Rudenko 1,  R.A. Ivanov 1,  A.B. Pshenichnikova 2,  and  V.I. Shvets 2

1  The BIOCAD Biopharmaceutical Co.,  198515, St.-Petersburg Russia

2  The Lomonosov Moscow State University for Fine Chemical Technology,  119571, Moscow Russia

e-mail:  korjavin@biocad.ru

 

A modification of recombinant human IFN beta-1a by polyethylene glycol at α-amino group of N-terminal methionine has been performed. The PEGylation was conducted using activated butyraldehyde PEG derivative with molecular mass of 30000 Da. As a result of the multifactorial experiment, the dependence of monoPEGylated protein yield on the reaction conditions was demonstrated, and optimal PEGylation conditions were established. We developed a one-step chromatographic purification scheme allowing to obtain the monoPEG-IFN beta-1a conjugate with the above 98% purity . Mass-spectrometric studies showed that the molecular mass of the conjugate is 54130 Da, wherein the N-terminal methionine is associated with one PEG molecule. The obtained PEG-IFN beta-1a has specific antiviral activity, and it can be considered as a perspective candidate to the design of long-acting medicine for the prolonged treatment of multiple sclerosis.

 

Key words:  human IFN-beta,  modification,  multiple sclerosis,  polyethylene glycol,  prolonged release form.

 

29.05.2015, 643 просмотра.

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