期刊文献+

肺炎链球菌荚膜多糖分子大小及分子量分析方法的比较 被引量:10

Comparison of methods for determination of molecular size and molecular weight of pneumococcal capsular polysaccharides
原文传递
导出
摘要 目的 3种方法分析10个血清型肺炎链球菌荚膜多糖(pneumococcal capsular polysaccharides,Pn Ps)样品的分子大小及分子量,并对这3种方法进行比较。方法采用Sepharose CL-4B凝胶排阻层析(SEC)技术检测1型、2型、4型、8型、9N型、12F型、14型、18C型、19A型和23F型Pn Ps各2~3批次样品在色谱柱中的分配系数(KD)和样品组分特征;采用高效凝胶排阻色谱-示差折光检测器(high performance size-exclusion chromatography with refractive index,HPSEC-RI)法建立Pullulan标准品分子量标准曲线,计算各Pn Ps样品的重均分子量(weight-average molecular weight,Mw);采用高效凝胶排阻色谱-多角度激光光散射仪/示差折光检测器联用技术(high performance size-exclu-sion chromatography with multi-angle laser light scattering and refractive index,HPSEC-MALLS/RI)检测各Pn Ps样品的Mw、数均分子量(number-average molecular weight,Mn)、多分散水平(Mw/Mn)及均方根旋转半径(root mean square ratio of gyration,Rg);对HPSEC-RI法和HPSEC-MALLS/RI法检测Pullulan分子量标准品的结果进行比较。结果 SEC技术不同化学方法检测的同批次Pn Ps样品的KD值十分接近,且主峰形态也比较近似,个别批次Pn Ps不同化学方法获得的峰形有差别。经HPSEC-RI检测Pullulan分子量标准品获得的回归方程为y=-0.278 x+10.13,r2=0.998,各Pn Ps样品的分子量为1.812×105~1.246×106。HPSEC-MALLS/RI联用技术检测的各批Pn Ps样品的Mw分布于9.898×105~1.471×105,Rg为84.9~25.4 nm,同一血清型不同批次样品Rg间的大小关系与其Mw间的大小关系具有较好一致性。结论结合多种方法检测多糖样品的分子大小和分子量,可更客观、全面地了解Pn Ps分子的各种性质。 Objective To compare three methods for determination of molecular size and molecular weight of pneumococcal capsular polysaccharides(Pn Ps). Methods Pn Ps of serotypes 1, 2, 4, 8, 9N, 12 F, 14, 18 C, 19 A and 23 F,2 ~ 3 batches for each, were determined for distribution coefficient(KD) value and composition specificity by sizeexclusion chromatography(SCE) with Sepharose CL-4B media. Standard molecular weight distribution curve was plotted by high performance size-exclusion chromatography-differential refractive index detector(HPSEC-RI) with Pullulan standard to calculate the weight-average molecular weight(Mw) of various Pn Ps samples. The Mw, number-average molecular weight(Mn), Mw/ Mnand root mean square ratio of gyration(Rg)of various Pn Ps samples were determined by HPSEC-mutiple angle laser light scattering apparatus(MALLS) / RI. The determination results of Pullulan standard by HPSEC-RI and HPSEC-MALLS / RI were compared. Results The KDvalues and shapes of major peaks of Pn Ps samples of the same batch determined by various methods were similar. However, the peak shapes of samples of individual batches determined by various methods were different. The regression equation of Pullalan standard determined by HPSEC-RI were y =-0. 278 x + 10. 13,r2 = 0. 998, while the molecular weights of various Pn Ps samples were 1. 812 × 105 ~ 1. 246 ×106. The Mwand Rgof Pn Ps samples of various batches determined by HPSEC-MALLS / RI were 9. 898 × 105~ 1. 471 ×105and 84. 9 ~ 25. 4 nm respectively. The Rgof samples of the same serotypes of various batches were consistent with their Mw. Conclusion Determination of molecular size and weight of Pn Ps samples by various methods was helpful to understanding the properties of Pn Ps molecules more objectively and more comprehensively.
出处 《中国生物制品学杂志》 CAS CSCD 2015年第9期947-955,960,共10页 Chinese Journal of Biologicals
关键词 肺炎链球菌荚膜多糖 分子大小 分子量 Pneumococcal capsular polysaccharides (PnPs) Molecular size Molecular weight
  • 相关文献

参考文献13

  • 1Morens DM,Taubenberger JK,Fauci AS.Predominant role of bacterial pneumonia as a cause of death in pandemic influenza:implications for pandemic influenza preparedness[J].J Infect Dis,2008,198(7):962-970.
  • 2韩菲,曹欣,胡鹏,张明华,郝倩,张美香,任涛,唐秀丽,何佳琦,王婷婷,郑海发,魏文进.23价肺炎链球菌荚膜多糖疫苗的研制[J].国际生物制品学杂志,2010,33(3):134-138. 被引量:5
  • 3Bednar B,Hennessey JP Jr.Molecular size analysis of capsular polysaccharide preparations from Streptococcus pneumonia[J].Carbohydr Res,1993,243(1):115-130.
  • 4World Health Organization.Recommendations to assure the quality,safety of pneumococcal conjugate vaccine[S].Geneva:WHO,2010.
  • 5柴雁菁,左智洁,马波,向左云,施競,周红军,徐东梅,黄镇.肺炎链球菌5型发酵工艺的优化[J].现代生物医学进展,2012,12(9):1660-1664. 被引量:4
  • 6European Directorate for the Quality of Medicines and Health Care.European Pharmacopoeia 7.0[S].Strasbourg:Council of Europe,2010.
  • 7Nagy DJ.Universal calibration in aqueous size exclusion chromatography with on-line differential viscometry using commercial TSK-PW columns[J].J Liq Chromatogr,1990,13(4):677-691.
  • 8Lesec J,Volet G.Data treatment in aqueous GPC with on-line viscometer and light scattering detectors[J].J Liq Chromatogr,1990,13(5):831-849.
  • 9于世林.高效液相色谱方法及应用[M].2版.北京:化学工业出版社,2010:157-159.
  • 10Mac Nair JE,Desai T,Teyral J,et al.Alignment of absolute and relative molecular size specifications for a polyvalent pneumococcal polysaccharide vaccine(PNEUMOVAX 23)[J].Biologicals,2005,33(1):49-58.

二级参考文献33

  • 1王剑虹,乔瑞洁,王欣茹,沈荣,赵萍.肺炎链球菌培养及其荚膜多糖的制备[J].甘肃科学学报,2004,16(3):40-44. 被引量:4
  • 2Pneumococcal conjugate vaccine for childhood immunization-WHO position paper[J].Wkly Epidem Rec,2007,82(7).93-104.
  • 3Fedson DS.Pneumococcal vaccine[M] // Plotkin SA,Morhmer EA.Vaccines.Philadelphia; WB Saun-ders Co,1988:271-299.
  • 4Robbing JB,Austrian R,Lee CJ,et al.Considerations for fomu-lating the second-generation pneumo-coccal capsular polysaccha-ride vaccine with emphasis on the cross-reactive types within groups[J].J Infect Dis,1983,148(6):1136-1159.
  • 5Directorate for the Quality of Medicines of the Council of Europe (EDQM).European Pharmacopoeia[M].5th ed.Strasbourg:EDQM,2004.
  • 6王博.中国跑步进入老龄社会[N].河北日报,2009-11-30(5).
  • 7Morris DL.Quantitative determination of carbohydrates with Drey-wood's anthrone reagent.Science,1984,107(2775):254-255.
  • 8李凡,谷鸿喜,黄敏.医学微生物学[M].教育出版社,2004.
  • 9秦绍明,余绍珍.实用传染病手册[M].军医出版社,1994.
  • 10Butler JC,Breiman RF,Campbell JF,et al.Pneumococcal polysaccharide,vaccine efficacy[J].JAMA,1993,270:1826-1831.

共引文献34

同被引文献70

引证文献10

二级引证文献24

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部