title: Screening for stability and compatibility conditions of recombinant human epidermal growth factor for parenteral formulation: effect of pH, buffers, and excipients.
abstract: A successful parenteral formulation can be developed by studying stability and compatibility of biopharmaceuticals as a function of solution composition. Here, we evaluate the influence of pH, buffers, ionic strength, protein concentration and presence of excipients on recombinant human epidermal growth factor (rhEGF) stability. The stability was accessed by reversed-phase high performance liquid chromatography (RP-HPLC), size exclusion chromatography (SEC-HPLC), enzyme-linked immunosorbent assay (ELISA), Far-UV circular dichroism (CD) and light scattering. The overall maximal stability was obtained in pH near to 7.0 in phosphate, Tris and histidine buffers as the results of the different methods revealed. The CD results revealed that this protein is stable in an extensive pH range. Aggregation of rhEGF was minimized at pH values ranged from 6.0 to 8.0 as indicated the SEC-HPLC and light scattering results. Nor the ionic strength neither the rhEGF concentration had significant effect on the reaction rate constants. Most rhEGF-excipient instability occurs among this protein and reducing sugars. Polymers like poly(ethylene glycol) (PEG) and polysorbates increased methionine oxidation. The rhEGF oxidation and deamidation were the most common degradation pathways. This research identified critical solution factors to be considered for the development of a successful rhEGF parenteral formulation.
authors.affiliation: Pharmaceutical Development Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box 6162, Havana 10600, Cuba. hector.santana@cigb.edu.cu
authors.forename: Héctor
authors.initials: H
authors.lastname: Santana
authors.forename: Yaima
authors.initials: Y
authors.lastname: González
authors.forename: Patricia Targon
authors.initials: PT
authors.lastname: Campana
authors.forename: Jesús
authors.initials: J
authors.lastname: Noda
authors.forename: Odalys
authors.initials: O
authors.lastname: Amarantes
authors.forename: Rosangela
authors.initials: R
authors.lastname: Itri
authors.forename: Alejandro
authors.initials: A
authors.lastname: Beldarraín
authors.forename: Rolando
authors.initials: R
authors.lastname: Páez
mesh_terms: D002021:Buffers; D004355:Drug Stability; D004815:Epidermal Growth Factor / Q000737:chemistry*; D005079:Excipients / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D010288:Parenteral Nutrition; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D002021:Buffers; D005079:Excipients; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor