Tuberculosis vaccine candidate: Characterization of H4-IC31 formulation and H4 antigen conformation
| Autor: | Federico Webster Magcalas, Sasmit S. Deshmukh, Kristen N. Kalbfleisch, Bruce Carpick, Marina Kirkitadze |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
medicine.medical_treatment Chemistry Pharmaceutical Recombinant Fusion Proteins Clinical Biochemistry Pharmaceutical Science Peptide Analytical Chemistry 03 medical and health sciences Antigen Adjuvants Immunologic Drug Discovery Zeta potential medicine Tuberculosis Particle Size Tuberculosis Vaccines Spectroscopy chemistry.chemical_classification Chemistry Spectrum Analysis Ligand (biochemistry) Fusion protein Protein tertiary structure 030104 developmental biology Biochemistry Oligodeoxyribonucleotides Tuberculosis vaccines Adjuvant |
| Zdroj: | Journal of pharmaceutical and biomedical analysis. 157 |
| ISSN: | 1873-264X |
| Popis: | Tuberculosis (TB) is one of the leading causes of death worldwide, making the development of effective TB vaccines a global priority. A TB vaccine consisting of a recombinant fusion protein, H4, combined with a novel synthetic cationic adjuvant, IC31®, is currently being developed. The H4 fusion protein consists of two immunogenic mycobacterial antigens, Ag85 B and TB10.4, and the IC31® adjuvant is a mixture of KLK, a leucine-rich peptide (KLKL5KLK), and the oligodeoxynucleotide ODN1a, a TLR9 ligand. However, efficient and robust methods for assessing these formulated components are lacking. Here, we developed and optimized phase analysis light scattering (PALS), electrical sensing zone (ESZ), and Raman, FTIR, and CD spectroscopy methods to characterize the H4-IC31 vaccine formulation. PALS-measured conductivity and zeta potential values could differentiate between the similarly sized particles of IC31® adjuvant and the H4-IC31 vaccine candidate and could thereby serve as a control during vaccine formulation. In addition, zeta potential is indicative of the adjuvant to antigen ratio which is the key in the immunomodulatory response of the vaccine. ESZ was used as an orthogonal method to measure IC31® and H4-IC31 particle sizes. Raman, FTIR, and CD spectroscopy revealed structural changes in H4 protein and IC31® adjuvant, inducing an increase in both the β-sheet and random coil content as a result of adsorption. Furthermore, nanoDSF showed changes in the tertiary structure of H4 protein as a result of adjuvantation to IC31®. Our findings demonstrate the applicability of biophysical methods to characterize vaccine components in the final H4-IC31 drug product without the requirement for desorption. |
| Databáze: | OpenAIRE |
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