Rapid Developability Assessments to Formulate Recombinant Protein Antigens as Stable, Low-Cost, Multi-Dose Vaccine Candidates: Case-Study With Non-Replicating Rotavirus (NRRV) Vaccine Antigens

• Published in: Journal of pharmaceutical sciences Vol. 110; no. 3; pp. 1042 - 1053
• Main Authors: Sawant, Nishant, Kaur, Kawaljit, Holland, David A., Hickey, John M., Agarwal, Sanjeev, Brady, Joseph R., Dalvie, Neil C., Tracey, Mary Kate, Velez-Suberbie, M. Lourdes, Morris, Stephen A., Jacob, Shaleem I., Bracewell, Daniel G., Mukhopadhyay, Tarit K., Love, Kerry R., Love, J. Christopher, Joshi, Sangeeta B., Volkin, David B.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2021; Elsevier
• Subjects: Antigens; Developability; Formulation; Multi-dose; Preservative; Recombinant Proteins; Rotavirus; Rotavirus Vaccines; Saccharomycetales; Stability; Vaccine; Stability; Multi-dose; Preservative; Rotavirus; Formulation; Developability; Vaccine
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1016/j.xphs.2020.11.039

A two-step developability assessment workflow is described to screen variants of recombinant protein antigens under various formulation conditions to rapidly identify stable, aluminum-adjuvanted, multi-dose vaccine candidates. For proof-of-concept, a series of sequence variants of the recombinant non-replicating rotavirus (NRRV) P[8] protein antigen (produced in Komagataella phaffii) were compared in terms of primary structure, post-translational modifications, antibody binding, conformational stability, relative solubility and preservative compatibility. Based on these results, promising P[8] variants were down-selected and the impact of key formulation conditions on storage stability was examined (e.g., presence or absence of the aluminum-adjuvant Alhydrogel and the preservative thimerosal) as measured by differential scanning calorimetry (DSC) and antibody binding assays. Good correlations between rapidly-generated developability screening data and storage stability profiles (12 weeks at various temperatures) were observed for aluminum-adsorbed P[8] antigens. These findings were extended and confirmed using variants of a second NRRV antigen, P[4]. These case-study results with P[8] and P[4] NRRV variants are discussed in terms of using this vaccine formulation developability workflow to better inform and optimize formulation design with a wide variety of recombinant protein antigens, with the long-term goal of rapidly and cost-efficiently identifying low-cost vaccine formulations for use in low and middle income countries.