TY - JOUR
T1 - Quality Improvement of Capsular Polysaccharide in Streptococcus pneumoniae by Purification Process Optimization
AU - Lee, Chankyu
AU - Chun, Hee Jin
AU - Park, Minchul
AU - Kim, Rock Ki
AU - Whang, Yoon Hee
AU - Choi, Seuk Keun
AU - Baik, Yeong Ok
AU - Park, Sung Soo
AU - Lee, Inhwan
N1 - Funding Information:
This study was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Family Welfare, South Korea.
Publisher Copyright:
© Copyright © 2020 Lee, Chun, Park, Kim, Whang, Choi, Baik, Park and Lee.
PY - 2020/2/4
Y1 - 2020/2/4
N2 - 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.
AB - 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.
KW - cell wall polysaccharide
KW - cetyltrimethylammonium bromide (CTAB) precipitation
KW - pneumococcal capsular polysaccharide
KW - purification
KW - vaccine
UR - http://www.scopus.com/inward/record.url?scp=85079612756&partnerID=8YFLogxK
U2 - 10.3389/fbioe.2020.00039
DO - 10.3389/fbioe.2020.00039
M3 - Article
AN - SCOPUS:85079612756
VL - 8
JO - Frontiers in Bioengineering and Biotechnology
JF - Frontiers in Bioengineering and Biotechnology
SN - 2296-4185
M1 - 39
ER -