Construction of a Simplex Search Framework for the Development of a New Specific and Robust Stability-Indicating Reversed-Phase High-Performance Liquid Chromatography Assay: Application for the Stability of a Peptide-Based Telomerase Vaccine.

In this work, a novel mathematical algorithm was described for the development of a high-performance liquid chromatography (HPLC) stability-indicating assay. This approach was applied for the stability study of a universal cancer peptide-based telomerase vaccine constituted of a mixture of two peptides. With only 18 experiments, the optimum HPLC gradient conditions were determined using a computer-assisted design of experiments-simplex algorithm software. In addition, the separation on the chromatogram of the isoaspartate peptide degradation product with the active pharmaceutical ingredients was achieved. This HPLC method was validated as per the International Committee on Harmonization guidelines. It was shown that this vaccine was physicochemical stable for 2 months when stored in borosilicate glass vials at -20°C and no formation of isoaspartate peptide was detected. In addition, this vaccine was stable for up to 10 h at 4°C or room temperature after thawing the samples. No impact of potential temperature excursion on the vaccine concentration in the borosilicate glass vial was observed.