BIOSIMILAR COMPARABILITY STUDIES

Kymos is a GMP-certified an EMA and FDA inspected CRO offering full biosimilar comparability studies. Releasing biosimilar drugs to the market is extremely challenging and calls for well-equipped laboratories able to conduct all assays, ensuring purity, identity and potency as well as GMP certifications and manufacturing authorizations, subject to periodic inspections by regulatory bodies such as the FDA and the EMA. We follow a step-by-step comparability model for biosimilar drugs focused on critical quality attributes (CQAs), identified and classified according to their risk ranking. The number and nature of required assays is determined from the list below on a case-by-case basis:

GENERAL ANALYTICAL TECHNIQUES	STRUCTURAL ANALYSIS
A preliminary overview of the quality attributes of the molecule during initial development phases: Electrophoresis (PAGE, SDS-PAGE, Bioanalyzer, IEF) Western Blot Extinction Coefficient, Bradford, Lowry, BCA (total protein quantitation) HPLC AccQ-Tag® (Waters) and OPA® (Agilent) for amino-acid analysis Sulfide quantitation by Ellman method Antibody isotyping	Full molecular identification for strain selection, bioprocess and final product characterization: Intact protein mass (Electrospray MS, MALDI-TOF) Peptide mapping, amino acid sequencing (LC-UV-MS/MS, QTof, QTRAP) N and C terminal sequencing of intact protein (MALDI-TOF and Edman degradation) Glycosylation and phosphorylation sites (LC-UV-MS) Glycosylation Profiles (GC, LC, MS/MS, capillary electrophoresis) Monosaccharide profile (LC and GC)
CONFORMATIONAL ANALYSIS	IDENTITY, CONTENT, PROTEIN IMPURITIES
Protein folding and 3D conformation determine biological activity and immunogenicity: Circular Dichroism (CD) Ultraviolet spectroscopy (UV) Fluorescence spectroscopy (FL) Infrared spectroscopy (FTIR)	Chromatographic patterns are quality indicators of the purification process: Liquid chromatographic patterns (RP-; SEC-; Ion Exchange-; and Affinity-HPLC/UPLC plus LC-UV-MS) Capillary electrophoresis (reduced and non-reduced) Imaged capillary isoelectric focusing (icIEF) ELISA, ECLA, RIA
PROCESS-RELATED IMPURITIES	BIOLOGICAL ACTIVITY
Other quality indicators of the purification process and also linked to safety concerns: Host Cell Proteins (specific ELISA) Chemical contaminants (HPLC, GC) Elemental impurities (AAS, ICP-MS) Nitrosamine impurities (LC-MS/MS, HS-GC-MS) DNA (qPCR and colorimetric commercial kits) Mycoplasma (PCR commercial kits) Endotoxins (colorimetric or gel-clot test) Bioburden	Biological tests to confirm molecule activity: Binding studies (ELISA, ECLA, RIA, SPR Biacore®) Potency assays (Cell –based assays) Competitive inhibition ELISA assays for vaccines