DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14

Zanardo, Rafaela Tais; Ferri, Anne Letícia Silva; Figueiredo, Douglas Borges De; Kraschowetz, Stefanie; Cabrera-Crespo, Joaquín; Gonçalves, Viviane Maimoni

doi:10.1590/0104-6632.20160333s20150140

Cited by 7 publications

(7 citation statements)

References 15 publications

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“…Other authors have added sodium deoxycholate after fermentation but before collecting cells and sourced CPS from the supernatant or filtrate (Macha et al, 2014 ). Zanardo and coworkers prefer to inactivate the bacteria with thimerosal and remove cells by filtration because lysing with detergent breaks down cell membranes, releasing large amounts of intracellular contaminants that make the purification process difficult (Zanardo et al, 2016 ). The main contaminants requiring separation include proteins, nucleic acids and cell-wall carbohydrates (C-Carbohydrates).…”

Section: Methods Of Purificationmentioning

confidence: 99%

“…Furthermore, Zanardo and coworkers evaluated a new purification process for serotype 14 (Zanardo et al, 2016 ), assaying three purification strategies consisting of the following steps: cell separation by tangential microfiltration, concentration by tangential ultrafiltration (50 kDa), diafiltration in the presence of sodium dodecyl sulfate using a 30-kDa ultrafiltration membrane, precipitation with 5% trichloroacetic acid, precipitation with 20 and 60% ethanol, and anion exchange chromatography. All processes achieved the required purity regarding nucleic acids (≤ 2%) and proteins (≤ 3%).…”

Section: Methods Of Purificationmentioning

confidence: 99%

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Purification of Capsular Polysaccharides of Streptococcus pneumoniae: Traditional and New Methods

Morais

Dee

Suárez

2018

Front. Bioeng. Biotechnol.

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Pneumonia caused by Streptococcus pneumoniae is a major bacterial disease responsible for many deaths worldwide each year and is particularly dangerous in children under 5 years old and adults over 50. The capsular polysaccharide (CPS) constitutes the outermost layer of the bacterial cell and is the main virulence factor. Regardless of whether pharmaceutical agents are composed of CPS alone or protein-conjugated CPS, CPS purification is essential for the development of vaccines against S. pneumoniae. These vaccines are effective and safe but remain quite expensive. This review describes the methods currently available for CPS purification. Advances in CPS purification methods are aimed at improvements in quality and yield and, above all, process simplification.

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Section: Methods Of Purificationmentioning

confidence: 99%

Section: Methods Of Purificationmentioning

confidence: 99%

Purification of Capsular Polysaccharides of Streptococcus pneumoniae: Traditional and New Methods

Morais

Dee

Suárez

2018

Front. Bioeng. Biotechnol.

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“…Several methods for purification of CPS from S. pneumoniae have been reported, and advances in technology have enabled production of CPS with high purity and yield (Suárez et al, 2001;Macha et al, 2014;Yuan et al, 2014;Zanardo et al, 2016;Morais et al, 2018). However, while these methods were optimized for specific serotypes of CPS such as type14 (Suárez et al, 2001;Zanardo et al, 2016), types 3, 6B, 14, 19F, and 23F (Macha et al, 2014), and types 1, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F (Yuan et al, 2014, our method can be universally applied to all 15 serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 11A, 14, 18C, 19A, 19F, 22F, and 23F) contained in a newly developing 15-valent conjugate vaccine, although there is one determination step whether to use supernatant or precipitant depending on the charge of the target CPS after CTAB precipitation. Methods mentioned above do not use traditional CTAB precipitation which is our main separation step, and we combined this with other techniques including acid precipitation, ethanol precipitation, and HA chromatography and this ensured high purity of CPS and low CWPS contamination.…”

Section: Discussionmentioning

confidence: 99%

Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization

Lee

Chun

Park

et al. 2020

Front. Bioeng. Biotechnol.

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Streptococcus pneumoniae is the causative agent of many diseases, most notably pneumonia. Most of the currently used vaccines to protect against this pathogen employ pneumococcal capsular polysaccharides (CPSs) as antigens, but purifying CPS of sufficient quality has been challenging. A purification process for CPS comprising conventional methods such as ultrafiltration, CTAB precipitation, and chromatography was previously established; however, this method resulted in high cell wall polysaccharide (CWPS) contamination, especially for serotype 5. Thus, a better purification method that yields CPS of a higher quality is needed for vaccine development. In this study, we significantly reduced CWPS contamination in serotype 5 CPS by improving the ultrafiltration and CTAB precipitation steps. Moreover, by applying an acid precipitation process to further remove other impurities, serotype 5 CPS was obtained with a lower impurity such as decreased nucleic acid contamination. This improved method was also successfully applied to 14 other serotypes (1,

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“…The nontagged recombinant fragment of PspA from clade 4 (PspA4Pro) and capsular polysaccharide from S. pneumoniae serotype 14 were produced and purified at Departamento de Engenharia Química (UFSCar) and Centro de Biotecnologia (Instituto Butantan) as described previously (36,37). Adipic acid dihydrazide (ADH) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) were from Sigma-Aldrich.…”

Section: Methodsmentioning

confidence: 99%

Conjugation of PspA4Pro with Capsular Streptococcus pneumoniae Polysaccharide Serotype 14 Does Not Reduce the Induction of Cross-Reactive Antibodies

Silva

Converso

Gonçalves

et al. 2017

Clin Vaccine Immunol

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Current pneumococcal vaccines are composed of bacterial polysaccharides as antigens, plain or conjugated to carrier proteins. While efficacious against vaccine serotypes, epidemiologic data show an increasing incidence of infections caused by nonvaccine serotypes of The use of pneumococcal surface protein A (PspA) as a carrier protein in a conjugate vaccine could help prevent serotype replacement by increasing vaccine coverage and reducing selective pressure of serotypes. PspA is present in all pneumococcal strains, is highly immunogenic, and is known to induce protective antibodies. Based on its sequence, PspA has been classified into three families and six clades. A PspA fragment derived from family 2, clade 4 (PspA4Pro), was shown to generate antibodies with a broad range of cross-reactivity, across clades and families. Here, PspA4Pro was modified and conjugated to capsular polysaccharide serotype 14 (PS14). We investigated the impact of conjugation on the immune response induced to PspA4Pro and PS14. Mice immunized with the PS14-mPspA4Pro conjugate produced higher titers of anti-PS14 antibodies than the animals that received coadministered antigens. The conjugate induced antibodies with opsonophagocytic activity against PS14-carrying strains, as well as against a panel of strains bearing PspAs from five clades (encompassing families 1 and 2) bearing a non-PS14 serotype. Furthermore, mice immunized with PS14-mPspA4Pro were protected against nasal colonization with a nonrelated strain bearing PspA from clade 1, serotype 6B. These results demonstrate that the cross-reactivity mediated by PspA4Pro is retained following conjugation, supporting the use of PspA4 as a carrier protein in order to enhance pneumococcal vaccine coverage and encourage its further investigation as a candidate in future vaccine designs.

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DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14

Cited by 7 publications

References 15 publications

Purification of Capsular Polysaccharides of Streptococcus pneumoniae: Traditional and New Methods

Purification of Capsular Polysaccharides of Streptococcus pneumoniae: Traditional and New Methods

Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization

Conjugation of PspA4Pro with Capsular Streptococcus pneumoniae Polysaccharide Serotype 14 Does Not Reduce the Induction of Cross-Reactive Antibodies

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