Optimization of Capsular Polysaccharide Production by Streptococcus pneumoniae Type 3

Jin, Sheng-De; Kim, Young‐Min; Kang, Hee-Kyoung; Jung, Seung-Jin; Kim, Doman

doi:10.4014/jmb.0903.3027

Cited by 7 publications

(2 citation statements)

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“…Various cell culture media for the growth of fastidious organisms such as S. pneumoniae are known. Most of the media used are available commercially and contain supplements such as animal extracts, yeast extract, or other sources of carbon and nitrogen, including sources of vegetable origin (Restrepo et al, 2005;Jin et al, 2009). These components enrich the nutritional quality of the basal media, to support the growth of the bacteria.…”

Section: Discussionmentioning

confidence: 99%

Economic Evaluation of <i>Streptococcus Pneumoniae</i> Culture Media

Morais¹,

Suárez²

2016

American Journal of Biochemistry and Biotechnology

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Streptococcus pneumoniae, a major bacterial pathogen worldwide, requires a complex medium for culture, which complicates harvesting of antigens for vaccine production. Here we present an economic evaluation of factors affecting the production cost of a chemically defined medium for the scale up of fermentation of S. pneumoniae, compared with the most traditional fermentation media. A basic laboratory-scale experiment of 1 L and an industrial-scale production of 100 L were assumed. The economic analysis results show that at 1 L lab-scale the Chemically Defined Medium (CDM) is the most expensive (US$35/L) compared to Tryptic Soy Broth medium (TSB) and Tryptic Soy Vegitone medium (TSBv),

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Section: Discussionmentioning

confidence: 99%

Economic Evaluation of <i>Streptococcus Pneumoniae</i> Culture Media

Morais¹,

Suárez²

2016

American Journal of Biochemistry and Biotechnology

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“…Almost all currently marketed glycoconjugate vaccines contain isolated, large and heterogeneous glycans 8 9 . These vaccines are very successful, but technically, the isolation from natural sources is challenging.…”

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confidence: 99%

Cross Reactive Material 197 glycoconjugate vaccines contain privileged conjugation sites

Möginger

Resemann

Martin

et al. 2016

Sci Rep

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Production of glycoconjugate vaccines involves the chemical conjugation of glycans to an immunogenic carrier protein such as Cross-Reactive-Material-197 (CRM197). Instead of using glycans from natural sources recent vaccine development has been focusing on the use of synthetically defined minimal epitopes. While the glycan is structurally defined, the attachment sites on the protein are not. Fully characterized conjugates and batch-to-batch comparisons are the key to eventually create completely defined conjugates. A variety of glycoconjugates consisting of CRM197 and synthetic oligosaccharide epitopes was characterised using mass spectrometry techniques. The primary structure was assessed by combining intact protein MALDI-TOF-MS, LC-MALDI-TOF-MS middle-down and LC-ESI-MS bottom-up approaches. The middle-down approach on CNBr cleaved glycopeptides provided almost complete sequence coverage, facilitating rapid batch-to-batch comparisons, resolving glycan loading and identification of side products. Regions close to the N- and C-termini were most efficiently conjugated.

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Identifying and optimization of critical process parameters for the modulation of polysaccharide molecular size in Streptococcus pneumoniae serotype-1

Sangareddy,

Mallu,

Matur

et al. 2024

Discov Appl Sci

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Maintaining the molecular size (MS) of Streptococcus pneumoniae capsular polysaccharide within specified range is essential for manufacture of conjugate vaccines, either through physical or acid hydrolysis before use in the conjugation process. Polysaccharide MS typically reduced, with high-pressure homogenization as an approach, for preserving their chemical structure. When the average MS of Pneumococcal capsular polysaccharide serotype-1(CPS1) exceeds 1200 kDa during fermentation, using a high-pressure homogenizer to reduce its MS to 150–250 kDa can become extremely difficult. Even after multiple homogenization cycles, obtaining polysaccharide of the required size can be challenging. Moreover, exceeding a certain number of homogenization cycles can negatively impact the stability, yield, and conjugation efficiency. To control polysaccharide MS, we conducted a design of experiments (DOE) study focused on the optimization of the fermentation process, employing serotype-1 as a represntative case. The successful optimization of these CPPs was achieved in a consistent and reproducible manner. Systematic evaluation by DOE based process optimization has provided valuable insights into precise polysaccharide manufacturing control of polysaccharide MS. Our findings confirm that maintaining Hy-Soy™ at 20-30 g/L and yeast extract at 1–3 g/L in the fermentation media, with a feed concentration of 2–3.5 g/L/H and 0.1–0.5 VVM of air, consistently yields polysaccharide with MS of < 1200 kDa. This strategy that can be extended to other S. pneumoniae serotype polysaccharide production. Graphical abstract

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Optimization of Capsular Polysaccharide Production by Streptococcus pneumoniae Type 3

Cited by 7 publications

References 0 publications

Economic Evaluation of <i>Streptococcus Pneumoniae</i> Culture Media

Economic Evaluation of <i>Streptococcus Pneumoniae</i> Culture Media

Cross Reactive Material 197 glycoconjugate vaccines contain privileged conjugation sites

Identifying and optimization of critical process parameters for the modulation of polysaccharide molecular size in Streptococcus pneumoniae serotype-1

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