国际生物制品学杂志
國際生物製品學雜誌
국제생물제품학잡지
INTERNATIONAL JOURNAL OF BIOLOGICALS
2014年
3期
114-117
,共4页
王伟%马雷钧%沈坚%陈晓怿%马相虎
王偉%馬雷鈞%瀋堅%陳曉懌%馬相虎
왕위%마뢰균%침견%진효역%마상호
嗜血菌,流感,b型%多糖类,细菌%纯化工艺
嗜血菌,流感,b型%多糖類,細菌%純化工藝
기혈균,류감,b형%다당류,세균%순화공예
Haemophilus influenzae,type b%Polysaccharides,bacterial%Purification process
目的 优化b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)多糖纯化工艺,替代传统的苯酚结合乙醇方法.方法 采用十六烷基三甲基溴化铵提取粗糖,以脱氧胆酸钠结合乙醇多级纯化进一步纯化粗糖,通过超滤浓缩和除菌过滤获得纯化的Hib多糖.采用优化的纯化工艺以中试和放大规模各纯化3批Hib多糖,按照《中华人民共和国药典》2010年版三部的要求检测纯化的Hib多糖.结果 中试和放大规模纯化的各3批Hib多糖的相对分子质量分别为621 800、634 400、659 900和597 200、612 100、583 400,均符合规定的标准.中试和放大规模纯化的各3批Hib多糖的细菌内毒素含量分别为2.0、0.5、0.7 EU/μg和4.0、2.0、1.0 EU/μg,均明显低于规定的标准.鉴别试验显示,中试和放大规模纯化的各批Hib多糖均可与标准Hib抗血清形成明显的沉淀线.结论 优化的Hib多糖纯化工艺具有较好的稳定性,且易于工艺放大,可替代传统工艺用于Hib多糖的纯化.
目的 優化b型流感嗜血桿菌(Haemophilus influenzae type b,Hib)多糖純化工藝,替代傳統的苯酚結閤乙醇方法.方法 採用十六烷基三甲基溴化銨提取粗糖,以脫氧膽痠鈉結閤乙醇多級純化進一步純化粗糖,通過超濾濃縮和除菌過濾穫得純化的Hib多糖.採用優化的純化工藝以中試和放大規模各純化3批Hib多糖,按照《中華人民共和國藥典》2010年版三部的要求檢測純化的Hib多糖.結果 中試和放大規模純化的各3批Hib多糖的相對分子質量分彆為621 800、634 400、659 900和597 200、612 100、583 400,均符閤規定的標準.中試和放大規模純化的各3批Hib多糖的細菌內毒素含量分彆為2.0、0.5、0.7 EU/μg和4.0、2.0、1.0 EU/μg,均明顯低于規定的標準.鑒彆試驗顯示,中試和放大規模純化的各批Hib多糖均可與標準Hib抗血清形成明顯的沉澱線.結論 優化的Hib多糖純化工藝具有較好的穩定性,且易于工藝放大,可替代傳統工藝用于Hib多糖的純化.
목적 우화b형류감기혈간균(Haemophilus influenzae type b,Hib)다당순화공예,체대전통적분분결합을순방법.방법 채용십륙완기삼갑기추화안제취조당,이탈양담산납결합을순다급순화진일보순화조당,통과초려농축화제균과려획득순화적Hib다당.채용우화적순화공예이중시화방대규모각순화3비Hib다당,안조《중화인민공화국약전》2010년판삼부적요구검측순화적Hib다당.결과 중시화방대규모순화적각3비Hib다당적상대분자질량분별위621 800、634 400、659 900화597 200、612 100、583 400,균부합규정적표준.중시화방대규모순화적각3비Hib다당적세균내독소함량분별위2.0、0.5、0.7 EU/μg화4.0、2.0、1.0 EU/μg,균명현저우규정적표준.감별시험현시,중시화방대규모순화적각비Hib다당균가여표준Hib항혈청형성명현적침정선.결론 우화적Hib다당순화공예구유교호적은정성,차역우공예방대,가체대전통공예용우Hib다당적순화.
Objective To optimize the purification process of Haemophilus influenzae type b (Hib) polysaccharide for replacing the traditional combination of phenol and ethanol.Methods Crude Hib polysaccharides were extracted with Cetavlon and further purified with the combination of sodium deoxycholate and ethanol multistage purification,and purified polysaccharides were obtained by ultrafiltration and sterilizing filtration.Three batches of purified Hib polysaccharides were prepared with the optimized purification precess in pilot and other 3 batches in amplifying scales.Prepared Hib polysaccharides were detected in accordance with the Chinese Pharmacopoeia 2010 edition (Volume Ⅲ).Results The relative molecular masses of each 3 batches of Hib polysaccharides purified in pilot and amplifying scales were 621 800,634 400,659 900 and 597 200,612 100,583 400,respectively,and all met standard requirement.The contents of bacterial endotoxin of each 3 batches of Hib polysaccharides purified in pilot and amplifying scales were 2.0,0.5,0.7 and 4.0,2.0,1.0,respectively,and were all significantly below standard requirement.The identity test showed every batch of Hib polysaccharides purified in pilot and amplifying scales formed a strong precipitin line with antiserum to Hib.Conclusion The optimized purification process is stable and easy to scale-up,and can replace the traditional purification process of Hib polysaccharide.
