Establecimiento de un método de cromatografía líquida de alta resolución para la cuantificación de proteína no enlazada en el conjugado de la vacuna SOBERANA®02
Year: 2026
Type: article
Abstract: SOBERANA®02 is a vaccine based on a protein subunit platform, produced by chemically conjugating the recombinant receptor-binding domain of the severe acute respiratory syndrome coronavirus 2 spike protein to tetanus toxoid absorbed in aluminum hydroxide. Quantifying the unbound protein in the conjugate is a critical attribute for evaluating the effectiveness of the conjugation process and ensuring the quality of the vaccine product. The objective of this work was to validate an HPLC method for quantifying the unbound protein, establishing an acceptance limit of less than 20 %. Three batches of conjugate were used and evaluated by size-exclusion chromatography on a system coupled to a diode array detector and a Superdex 75 column with a 10/300 increment (Cytiva). Concentrations were determined from a calibration curve using a reference material, and the following parameters were evaluated: system suitability (linearity and range), precision, accuracy, specificity, limit of quantification, and limit of detection. The coefficient of variation for evaluating system suitability in terms of retention time precision was 0.011 %, and the skewness factor was 1.18. The coefficients of variation in the intra- and inter-assay precision tests were within the established ranges for each (≤ 1 % and ≤ 5 %). The accuracy of the method was demonstrated by the recovery percentages obtained (90 % and 110 %), and at all concentration levels studied, a coefficient of variation of < 2 % was obtained between recoveries. The calculated limit of quantification for the method was 0.0129 µg/mL, and the limit of detection was 0.0043 µg/mL. The results obtained confirm the robustness of the method and support its use for the quantification of unbound proteins, thus ensuring the quality of the conjugate in vaccine samples.
Source: Vaccimonitor/VacciMonitor
Institution: Finlay Institute
Language: Spanish
Subfield: Analytical Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: diamond
OpenAlex ID: W7224979786