国际生物制品学杂志
國際生物製品學雜誌
국제생물제품학잡지
INTERNATIONAL JOURNAL OF BIOLOGICALS
2013年
4期
177-181
,共5页
张霖阳%潘殊男%李世慧%徐鹤%周峻凯%肖詹蓉
張霖暘%潘殊男%李世慧%徐鶴%週峻凱%肖詹蓉
장림양%반수남%리세혜%서학%주준개%초첨용
CHO细胞%百日咳毒素%方法
CHO細胞%百日咳毒素%方法
CHO세포%백일해독소%방법
CHO cells%Pertussis toxin%Method
目的 建立检测百日咳毒素(pertussis toxin,PT)活性的中国仓鼠卵巢(Chinese hamster ovary,CHO)细胞簇集法,并将建立的方法应用于无细胞百日咳疫苗生产的质量控制.方法 将PT标准品系列稀释后与培养的CHO细胞混合,观察PT引起CHO细胞簇集的敏感度,并研究反应时间、反应溶液的pH值、缓冲液浓度、加样顺序等因素对PT引起CHO细胞簇集的影响.应用建立的方法检测百日咳疫苗生产过程中脱毒前后的PT活性及丝状血凝素和百日咳黏着素组分中残留的PT活性.结果 PT使CHO细胞发生最佳簇集的理想反应时间是24h,最适pH值是7～8,适宜氯化钠缓冲液浓度为0.15 mol/L,最优加样顺序是先加CHO细胞再加PT.建立的CHO细胞簇集法的检测敏感度为10pg/ml PT,且重复性良好.结论 CHO细胞簇集法可用于PT活性检测.
目的 建立檢測百日咳毒素(pertussis toxin,PT)活性的中國倉鼠卵巢(Chinese hamster ovary,CHO)細胞簇集法,併將建立的方法應用于無細胞百日咳疫苗生產的質量控製.方法 將PT標準品繫列稀釋後與培養的CHO細胞混閤,觀察PT引起CHO細胞簇集的敏感度,併研究反應時間、反應溶液的pH值、緩遲液濃度、加樣順序等因素對PT引起CHO細胞簇集的影響.應用建立的方法檢測百日咳疫苗生產過程中脫毒前後的PT活性及絲狀血凝素和百日咳黏著素組分中殘留的PT活性.結果 PT使CHO細胞髮生最佳簇集的理想反應時間是24h,最適pH值是7～8,適宜氯化鈉緩遲液濃度為0.15 mol/L,最優加樣順序是先加CHO細胞再加PT.建立的CHO細胞簇集法的檢測敏感度為10pg/ml PT,且重複性良好.結論 CHO細胞簇集法可用于PT活性檢測.
목적 건립검측백일해독소(pertussis toxin,PT)활성적중국창서란소(Chinese hamster ovary,CHO)세포족집법,병장건립적방법응용우무세포백일해역묘생산적질량공제.방법 장PT표준품계렬희석후여배양적CHO세포혼합,관찰PT인기CHO세포족집적민감도,병연구반응시간、반응용액적pH치、완충액농도、가양순서등인소대PT인기CHO세포족집적영향.응용건립적방법검측백일해역묘생산과정중탈독전후적PT활성급사상혈응소화백일해점착소조분중잔류적PT활성.결과 PT사CHO세포발생최가족집적이상반응시간시24h,최괄pH치시7～8,괄의록화납완충액농도위0.15 mol/L,최우가양순서시선가CHO세포재가PT.건립적CHO세포족집법적검측민감도위10pg/ml PT,차중복성량호.결론 CHO세포족집법가용우PT활성검측.
Objective To establish Chinese hamster ovary (CHO) cell clustering method for determination of activity of pertussis toxin (PT) and apply the established method to quality control in production process of acellular pertussis vaccine (APV).Methods PT standards diluted serially were mixed with CHO cells.Sensitivity of CHO cell clustering by induced PT was observed,and different influence factors on CHO cell clustering,including reaction time,pH value of reaction solution,buffer concentration and order of adding sample were studied.The established method was applied to PT activity detection before and after detoxification and residual PT activity in filamentous hemagglutinin and pertactin components in production process of APV.Results The optimal reaction time,pH value of reaction solution,buffer concentration and order of adding sample for the best CHO cell clustering by induced PT were 24 h,pH 7-8,0.15 mol/L,first adding CHO cells and then PT,respectively.Sensitivity of the established CHO cell clustering method was 10 pg/ml PT.This method had good repeatability.Conclusion The established CHO cell clustering method can be applied to PT activity detection.
