Ponente
Descripción
Abstract.
Background. For pharmaceutical proteins and peptides, the verification of the N-terminal sequenece is requested by regulatory authorities according to the ICHQ-6B Guideline for developing well-characterized pharmaceutical products. This information shold be used throughout all stages of drug development to demostrate comparability and consistency of productive batches. The N-terminal sequencing by stepwise Edmand degradation was established for CIGB-814, a synthetic peptide with anti-inflammatory properties that is currently being used in the treatment of severe and critically ill COVID-19 patients. Materials and Methods. Two-hundred pmol of the CIGB-814 was applied on a PVDF membrane, treated with methanol, and flushed with a N2 until complete dryness. The membrane was washed with ethanol 10% during 2 minutes, re-dried with N2 and it was introduced in the reactor of a Protein Sequencer PPSQ51A (Shimadzu, Japan); where 16 or more reaction cycles were scheduled. Results. The initial and repetitive yields were ≥50% and ≥80%, respectively considering Ala, Leu, and Val amino acids as recommended by the manufacter. The correlation coeficient was ≥0.90 for these parameters. Conclusions. Batches produced at the research level (Synthetic peptides laboratory at the CIGB) and at a industrial scale (in three different facilities) were characterized and in all cases a unique N-terminal sequence of fifteen amino acids was obtained in a full agreement with the expected for CIGB-814.
Key Words: Edmand Degradation, drug development, N-terminal sequencing, productive batches.
