Pegylated Recombinant Human GCSF (filgrastim)

Pegfilgrastim is a PEGylated form of the recombinant human granulocyte colony-stimulating factor (GCSF) analog filgrastim. It plays a crucial role in stimulating the production of white blood cells (neutrophils).
Pegfilgrastim’s half-life is significantly longer than the non-pegylated version, making it a valuable asset in medical practice.

Transfer of the end-to-end technology available in GMP-like environment.

mPEG20K-propionaldehyde (Pegylation Reagent) is prepared starting from commercially available raw mPEG20K-OH material, which is checked for purity
and quality by RPHPLC-Evaporative Light Scattering Detection (RPHPLC-ELSD) and
SDS-PAGE (Barium/iodine staining). Safety measures for proper handling and
disposal of the reagents involved are carefully checked.
The synthetic reaction batch is performed using 10 grams of starting mPEG20K-OH. The synthesis crude product is checked by RP-HPLC-ELSD after a test chemical reaction, in order to evaluate the aldehyde activity, which is at least 80%.
rhG-CSF is reacted with Pegylation Reagent in controlled conditions of buffer composition, protein concentration, reagent molar excess, pH, temperature and time. At the endpoint the reaction contains the monopegylated rhG-CSF (mPEG-rhG-CSF),
native rhG-CSF and oligomeric forms of pegylated rhGCSF. The reaction is analyzed by analytical RP-HPLC-ELSD to evaluate the yield of mPEG-rhG-CSF in the Pegylation Reaction. The batch size of the transfer can be up to the equivalent of
100 doses of mPEG-rhG-CSF.
mPEG-rhG-CSF is purified by chromatography. The purity of the product is at least
97% by SDS-PAGE (stained by Coomassie to visualize the protein portion and by
Barium/iodine to visualize the mPEG portion) and RP-HPLC analysis. Final yield of
the process is at least 40% of the rhG-CSF used in the pegylation reaction.

Pegylated rhG-CSF is used of severe myelosuppression abe neutropenia caused by a variety of conditions such as radiation exposure and chemotherapy for cancer treatment.
Pegylated rhG-CSF is also used in the research of hematopoietic stem and progenitor cells. It plays a crucial role in the sourcing, isolation, expansion, and differentiation of these cells.