Luis Mauricio

Structural Comparability of Biopharmaceuticals

Luís Maurício T. R. Lima This e-mail address is being protected from spam bots, you need JavaScript enabled to view it

Faculdade de Farmácia, Universidade Federal do Rio de Janeiro

Lab. for Pharmaceutical Biotechnology, Av. Carlos Chagas Filho 373, CCS, Bss24, Ilha do Fundão, RJ, RJ, 21941-902, Brazil

The efficacy and safety of biotechnological and biopharmaceuticals products can be largely affected by changes in folding and quaternary assembly of their biological active pharmaceutical ingredients (API). In the main pharmacopeias and in the ICH guidelines the structural analysis of biopharmaceuticals is limited to indirect evidences by assays such as proteolytic fingerprint and SEC analysis for high molecular weight oligomers. The combined use of mass spectroscopy (MS) and techniques for high resolution structural elucidation of macromolecules such as X-ray crystallography and nuclear magnetic resonance (NMR) would assist in the characterization of the API in the final, formulated product. To address this issue, we have subjected therapeutic proteins direct from final formulated products to structural analysis direct from commercial formulation using PX, NMR, MS, as well as oligomeric evaluation by small angle X-ray scattering (SAXS). We have extensively evaluated several validation parameters, by testing different batches, protein variants / analogues, biological API production methods, cell strains used in production (e.g., bacterial, yeast, combined biological and synthetic routes), NMR magnets, crystallization variables (such as buffers, precipitants, pH and temperature), X-ray sources and diffraction data collection strategies. We will discuss the use of MS, NMR fingerprint and PX in the comparability exercise of biological products subject to changes in their manufacturing process (ICH Q5E) and in the comparability track for the registration of biological products in Brazil (CP 49/2010), as well as a routine protocol for the precise chemical and structural characterization and proof of folding identity of biopharmaceuticals in the final, formulated product.

Key words: Biopharmaceutical; NMR; protein crystallography

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