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Puchkov I.A.,Russian Pharmaceutical Company CC Masterclone | Kononova N.V.,Russian Pharmaceutical Company CC Masterclone | Bobruskin A.I.,Russian Pharmaceutical Company CC Masterclone | Bairamashvili D.I.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | And 2 more authors.
Russian Journal of Bioorganic Chemistry | Year: 2012

With the purpose of creating an active prolonged-release pharmaceutical substance, modification of the recombinant human granulocyte colony-stimulating factor G-CSF (filgrastim) with polyethylene glycol (PEG, molecular mass 21.5 kDa) has been performed. The method for the preparation of the filgrastim PEG derivative intended to develop and scale-up the technological manufacturing process is described. Protein modification with PEG was performed by selective covalent attachment of the α-methyl-PEG-propionaldehyde molecule to the α-amino group of the N-terminal of the methionine amino acid residue of the recombinant G-CSF. The selected reaction conditions provide no less than 85% product yield of the total protein, a high protein concentration in the reaction mixture (more than 9 mg/mL) and allow us to reduce PEG consumption on the protein terminal α-amino group basis. RP HPLC and MALDI mass spectrometry data demonstrate that the preparation is modified by PEG at the N-terminal residue and contains no more than 10% of products with the higher degree of modification. © Pleiades Publishing, Ltd., 2012.

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