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Optimization of Fermentation Conditions Using Design of Experiment (DoE) to Enhance the Production of Antigenic

Amol Dattatraya Mahajan1,2, Pramod Maruti Jadhav2, Avinash Rohidas Memane2

  • 1MIT School of Bioengineering Sciences & Research, MIT Art Design and Technology University, Pune, India.

Biotechnology and Applied Biochemistry
|March 24, 2026
PubMed

Insights

Optimizing Streptococcus pneumoniae fermentation using Design of Experiments (DoE) significantly boosted vaccine antigen yield. This data-driven approach enhances scalability for pneumococcal vaccines, addressing global health needs.

Area of Science:

  • Biotechnology
  • Microbiology
  • Vaccine Development

Background:

  • Pneumococcal pneumonia, caused by Streptococcus pneumoniae, is a significant global health concern, particularly for young children and the elderly.
  • There is a critical need for scalable and high-yield production methods for Streptococcus pneumoniae vaccines.

Purpose of the Study:

  • To develop an innovative and optimized fermentation platform for Streptococcus pneumoniae (serotype 18C) production.
  • To enhance the yield of antigenic capsular polysaccharide through a data-driven approach.

Main Methods:

  • Utilized a data-driven Design of Experiment (DoE) approach, specifically Plackett-Burman design (PBD), to optimize media composition and fermentation parameters.
  • Investigated the effects of dextrose, MgSO4, pH, and aeration on bacterial growth and polysaccharide yield.
  • Employed downstream processing including centrifugation and diafiltration (100 kDa) for product recovery and purification.

Main Results:

  • Identified pH 7.1 as optimal for significantly enhancing antigenic capsular polysaccharide yield.
  • ANOVA analysis revealed a synergistic effect between dextrose and MgSO4 concentrations.
  • Lower aeration levels (0.2 vvm) were found to favor bacterial growth, consistent with the microaerophilic nature of S. pneumoniae.

Conclusions:

  • The optimized fermentation platform using PBD sets a new benchmark for pneumococcal fermentation, improving vaccine antigen yield and addressing scalability challenges.
  • Precise modulation of fermentation conditions is key to enhancing vaccine antigen production for industrial manufacturing.
  • This adaptive, data-driven bioprocess innovation paves the way for next-generation pneumococcal vaccine development.

Keywords:
Plackett–Burman design (PBD)Streptococcus pneumoniaeantigenic capsular polysaccharidefermentation optimizationmedia optimizationstatistical modelingvaccine manufacturing

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