التفاصيل البيبلوغرافية
| العنوان: |
Mechanism of Thimerosal-Induced Structural Destabilization of a Recombinant Rotavirus P[4] Protein Antigen Formulated as a Multi-Dose Vaccine. |
| المؤلفون: |
Kaur, Kawaljit1 (AUTHOR), Xiong, Jian1 (AUTHOR), Sawant, Nishant1 (AUTHOR), Agarwal, Sanjeev1 (AUTHOR), Hickey, John M.1 (AUTHOR), Holland, David A.1 (AUTHOR), Mukhopadhyay, Tarit K.1,2 (AUTHOR), Brady, Joseph R.3 (AUTHOR), Dalvie, Neil C.3 (AUTHOR), Tracey, Mary Kate3 (AUTHOR), Love, Kerry R.3 (AUTHOR), Love, J. Christopher3 (AUTHOR), Weis, David D.4 (AUTHOR), Joshi, Sangeeta B.1 (AUTHOR), Volkin, David B.1 (AUTHOR) volkin@ku.edu |
| المصدر: |
Journal of Pharmaceutical Sciences. Mar2021, Vol. 110 Issue 3, p1054-1066. 13p. |
| مصطلحات موضوعية: |
*ROTAVIRUSES, *COORDINATE covalent bond, *ANTIGENS, *VACCINES, *CYTOSKELETAL proteins |
| مستخلص: |
In a companion paper, a two-step developability assessment is presented to rapidly evaluate low-cost formulations (multi-dose, aluminum-adjuvanted) for new subunit vaccine candidates. As a case study, a non-replicating rotavirus (NRRV) recombinant protein antigen P[4] was found to be destabilized by the vaccine preservative thimerosal, and this effect was mitigated by modification of the free cysteine (C173S). In this work, the mechanism(s) of thimerosal-P[4] protein interactions, along with subsequent effects on the P[4] protein's structural integrity, are determined. Reversible complexation of ethylmercury, a thimerosal degradation byproduct, with the single cysteine residue of P[4] protein is demonstrated by intact protein mass analysis and biophysical studies. A working mechanism involving a reversible S-Hg coordinate bond is presented based on the literature. This reaction increased the local backbone flexibility of P[4] within the helical region surrounding the cysteine residue and then caused more global destabilization, both as detected by HX-MS. These effects correlate with changes in antibody-P[4] binding parameters and alterations in P[4] conformational stability due to C173S modification. Epitope mapping by HX-MS demonstrated involvement of the same cysteine-containing helical region of P[4] in antibody-antigen binding. Future formulation challenges to develop low-cost, multi-dose formulations for new recombinant protein vaccine candidates are discussed. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
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