Monday, July 27, 2009
P26
Simplified Purification Process for Capsular Polysaccharide from Streptococcus pneumoniae
Seung-Jin Jung, Interdisciplinary Program of Graduate School for Bioenergy & Biomaterials, 300 Yongbong-dong, Gwangju, South Korea, Sheng-De Jin, School of Biological Sciences and Technology, Chonnam National University, 300 Yongbong-dong, Gwangju, 500-757, South Korea, Young-Min Kim, School of Biological Sciences and Technology, The second stage of BK21, Korea Ministry of Education, The Research Institute for, Chonnam National University, 300 Yongbong-dong, Gwangju, 500-757, South Korea, and Doman Kim, School of Biological Sciences and Technology, Interdisciplinary Program of Graduate School for Bioenergy & Biomaterials, Chonnam National University, 300 Yongbong-dong, Gwangju, 500-757, South Korea.
Streptococcus pneumoniae is a major cause of invasive infection in young infants and older adults. Capsular polysaccharide (CPS) produced by S. pneumoniae is a virulence factor. CPS has also been used as a vaccine. On the basis of difference in CPS structure, S. pneumoniae can be divided into more than 90 serotypes. Among them 23 serotypes (1, 2, 3, 4, 5, 6b, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F and 33F) were responsible for about 90% of invasive disease.
Purification of CPS is complicate process and is required many kinds of reagents and steps. In this study, we simplified the purification process of CPS from Streptococcus pneumoniae serotype 19A by using ethanol fractionation and pH adjustment as following: First, a precipitant was removed from the cultured broth by ethanol fractionation (30~50%, v/v) method. The precipitant prepared with less than 30% and more than 50% of ethanol contained most contaminated proteins. Precipitation with 30% ethanol was performed at pH 9.5 after NaOH addition, and 50% ethanol precipitation was performed at neutral pH at 7.0. Small carbohydrates were easily removed by dialysis method with dH2O. By using simplified process from 1 L Brain Heart Infusion (BHI) culture the CPS obtained showed more than 95% purity.
Purification of CPS is complicate process and is required many kinds of reagents and steps. In this study, we simplified the purification process of CPS from Streptococcus pneumoniae serotype 19A by using ethanol fractionation and pH adjustment as following: First, a precipitant was removed from the cultured broth by ethanol fractionation (30~50%, v/v) method. The precipitant prepared with less than 30% and more than 50% of ethanol contained most contaminated proteins. Precipitation with 30% ethanol was performed at pH 9.5 after NaOH addition, and 50% ethanol precipitation was performed at neutral pH at 7.0. Small carbohydrates were easily removed by dialysis method with dH2O. By using simplified process from 1 L Brain Heart Infusion (BHI) culture the CPS obtained showed more than 95% purity.
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See more of The Annual Meeting and Exhibition 2009 (July 26 - 30, 2009)