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Published on: February 22, 2019
Evaluating the Compatibility of New Recombinant Protein Antigens (Trivalent NRRV) with a Mock Pentavalent Combination
Prashant Kumar1, David A Holland1, Kathryn Secrist1
1Department of Pharmaceutical Chemistry, Vaccine Analytics and Formulation Center, University of Kansas, Lawrence, KS 66047, USA.
Insights
Developing new combination vaccines for children in low-income countries faces challenges. Combining rotavirus antigens with pentavalent vaccines caused antigen instability due to adjuvants and preservatives.
Area of Science:
- Vaccinology
- Protein Chemistry
- Analytical Chemistry
Background:
- Combination vaccines improve pediatric immunization coverage and affordability, particularly in low- and middle-income countries (LMICs).
- Recombinant non-replicating rotavirus vaccine (NRRV) antigens present unique formulation challenges when combined with existing vaccine platforms.
Purpose of the Study:
- To investigate the analytical and formulation challenges of combining three recombinant non-replicating rotavirus vaccine (t-NRRV) antigens with a mock multidose pediatric pentavalent vaccine.
- To identify factors contributing to antigen instability and degradation in complex vaccine formulations.
Main Methods:
- Development of selective, stability-indicating competitive immunoassays for monitoring t-NRRV antigen binding to Alhydrogel (AH).
- Utilized a mouse immunogenicity assay for whole-cell pertussis (wP) antigen analysis.
- Evaluated antigen stability in mock pentavalent formulations containing various adjuvants (AH, Adju-Phos (AP)) and a preservative (thimerosal (TH)).
Main Results:
- Simple mixing of t-NRRV antigens with the pentavalent formulation led to antigen desorption from AH and increased degradation.
- Specific antigens, AP, and TH were identified as causes of deleterious effects on t-NRRV antigens.
- An AH-only pentavalent formulation reduced t-NRRV antigen desorption, but Hib antigen instability persisted.
- A consistent rank-ordering of t-NRRV antigen stability (P[8] > P[4] > P[6]) was observed across formulations.
Conclusions:
- Multidose combination vaccine formulations require careful consideration of antigen-adjuvant-preservative interactions.
- Addressing antigen stability issues is crucial for the successful development of new pediatric combination vaccines for LMICs.
- Lessons learned provide insights for future development of stable, multidose combination vaccines.
Abstract:
Introducing new recombinant protein antigens to existing pediatric combination vaccines is important in improving coverage and affordability, especially in low- and middle-income countries (LMICs). This case-study highlights the analytical and formulation challenges encountered with three recombinant non-replicating rotavirus vaccine (NRRV) antigens (t-NRRV formulated with Alhydrogel® adjuvant, AH) combined with a mock multidose formulation of a pediatric pentavalent vaccine used in LMICs. This complex formulation contained (1) vaccine antigens (i.e., whole-cell pertussis (wP), diphtheria (D), tetanus (T), Haemophilus influenza (Hib), and hepatitis B (HepB), (2) a mixture of aluminum-salt adjuvants (AH and Adju-Phos®, AP), and (3) a preservative (thimerosal, TH). Selective, stability-indicating competitive immunoassays were developed to monitor binding of specific mAbs to each antigen, except wP which required the setup of a mouse immunogenicity assay. Simple mixing led to the desorption of t-NRRV antigens from AH and increased degradation during storage. These deleterious effects were caused by specific antigens, AP, and TH. An AH-only pentavalent formulation mitigated t-NRRV antigen desorption; however, the Hib antigen displayed previously reported AH-induced instability. The same rank-ordering of t-NRRV antigen stability (P[8] > P[4] > P[6]) was observed in mock pentavalent formulations and with various preservatives. The lessons learned are discussed to enable future multidose, combination vaccine formulation development with new vaccine candidates.
