Pegylation of Human Interferon Beta 1A

Pegylation improves the therapeutic efficacy of the drug. Human Interferon Beta 1A is a protein with antiviral properties, used primarily in the treatment of multiple sclerosis. Pegylation enhances its stability, reduces its immunogenicity, and prolongs its half-life, thereby reducing the frequency of administration and potentially improving patient compliance.

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

mPEG20K-methyl-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 (RP-HPLC-
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 crude product of the synthesis is checked by RP-HPLC-ELSD after a test chemical reaction, in order to evaluate the aldehyde activity, which is at least 80%.
Protein Pegylation: 50 mg of IFNβ1a (provided by the company) in 10mM NaOAc - 5% sorbitol pH4.4 is concentrated by ultrafiltration (MWCO 10KDa) and reacted with mPEG20kDa-O-2-methylpropionaldehyde produced in house. The Recombinant human
Interferon Beta1a (IFN-Beta1a) with purity according to European Pharmacopoeia
is checked by RP-HPLC. IFN-Beta1a is then 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 IFN-Beta1a (mPEG-IFN-Beta1a), native IFN-Beta1a and oligomeric forms of pegylated IFN-Beta1a. The reaction is analyzed by analytical RP-HPLC-ELSD to evaluate the yield of mPEG-IFN-Beta1a in the Pegylation Reaction.
The mPEG-IFN-Beta1a is purified by chromatography. The purity of the product is at least 95% by SDS-PAGE (stained by Coomassie to visualize the protein portion and by Barium/iodine to visualize the mPEG portion) and RP-HPLC analysis.

Antiviral and Immunomodulatory Activities: IFN-β1A is secreted by cells in response to viral infections. It has antiviral, anti-proliferative, and multifunctional immunomodulatory activities.