Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization
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...
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Frontiers Media S.A.
2020-02-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fbioe.2020.00039/full |
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| author | Chankyu Lee Chankyu Lee Hee Jin Chun Minchul Park Rock Ki Kim Yoon Hee Whang Seuk Keun Choi Yeong Ok Baik Sung Soo Park Inhwan Lee |
| author_facet | Chankyu Lee Chankyu Lee Hee Jin Chun Minchul Park Rock Ki Kim Yoon Hee Whang Seuk Keun Choi Yeong Ok Baik Sung Soo Park Inhwan Lee |
| author_sort | Chankyu Lee |
| collection | DOAJ |
| description | 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, 3, 4, 6A, 6B, 7F, 9V, 11A, 14, 18C, 19A, 19F, 22F, and 23F). To assess the immunogenicity of the CPS from the 15 serotypes, two sets of 15-valent pneumococcal conjugate vaccines were prepared using the previous purification method and the improved method developed here; these vaccines were administered to a rabbit model. Enzyme-linked immunosorbent assay and opsonophagocytic assay demonstrated higher immunogenicity of the conjugate vaccine prepared using CPS produced by the improved purification process. |
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| format | Article |
| id | doaj.art-64040765039e46f594eae47f97441515 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-13T14:20:55Z |
| publishDate | 2020-02-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-64040765039e46f594eae47f974415152022-12-22T02:43:28ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-02-01810.3389/fbioe.2020.00039498353Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process OptimizationChankyu Lee0Chankyu Lee1Hee Jin Chun2Minchul Park3Rock Ki Kim4Yoon Hee Whang5Seuk Keun Choi6Yeong Ok Baik7Sung Soo Park8Inhwan Lee9R&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaDepartment of Biotechnology, Korea University, Seoul, South KoreaR&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaR&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaR&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaR&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaR&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaR&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaDepartment of Biotechnology, Korea University, Seoul, South KoreaR&D Center, EuBiologics Co., Ltd., Chuncheon-si, South KoreaStreptococcus 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, 3, 4, 6A, 6B, 7F, 9V, 11A, 14, 18C, 19A, 19F, 22F, and 23F). To assess the immunogenicity of the CPS from the 15 serotypes, two sets of 15-valent pneumococcal conjugate vaccines were prepared using the previous purification method and the improved method developed here; these vaccines were administered to a rabbit model. Enzyme-linked immunosorbent assay and opsonophagocytic assay demonstrated higher immunogenicity of the conjugate vaccine prepared using CPS produced by the improved purification process.https://www.frontiersin.org/article/10.3389/fbioe.2020.00039/fullpneumococcal capsular polysaccharidepurificationcetyltrimethylammonium bromide (CTAB) precipitationcell wall polysaccharidevaccine |
| spellingShingle | Chankyu Lee Chankyu Lee Hee Jin Chun Minchul Park Rock Ki Kim Yoon Hee Whang Seuk Keun Choi Yeong Ok Baik Sung Soo Park Inhwan Lee Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization Frontiers in Bioengineering and Biotechnology pneumococcal capsular polysaccharide purification cetyltrimethylammonium bromide (CTAB) precipitation cell wall polysaccharide vaccine |
| title | Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization |
| title_full | Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization |
| title_fullStr | Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization |
| title_full_unstemmed | Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization |
| title_short | Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization |
| title_sort | quality improvement of capsular polysaccharide in streptococcus pneumoniae by purification process optimization |
| topic | pneumococcal capsular polysaccharide purification cetyltrimethylammonium bromide (CTAB) precipitation cell wall polysaccharide vaccine |
| url | https://www.frontiersin.org/article/10.3389/fbioe.2020.00039/full |
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