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
6期
300-303
,共4页
α1抗胰蛋白酶%胰蛋白酶%发色底物法
α1抗胰蛋白酶%胰蛋白酶%髮色底物法
α1항이단백매%이단백매%발색저물법
Alpha 1-antitrypsin%Trypsin%Chromogenic substrate method
目的 验证优化的α1-抗胰蛋白酶(α1-antitrypsin,α1-AT)生物活性测定方法.方法 用发色底物法测定α1-AT活性,以牛胰蛋白酶替换冻干正常人血清来优化先前建立的方法,验证优化方法的准确性、重复性,并观察不同物质对优化方法的影响.结果 优化方法在胰蛋白酶质量浓度为0.01～0.05 g/L时线性关系良好,相关系数＞0.99.优化方法具有良好的准确性和重复性,当采用优化方法测定质量浓度为0.01、0.03、0.05 g/L胰蛋白酶时,胰蛋白酶的回收率分别为99.33％、94.44％和92.67％,变异系数分别为3.79％、1.66％和1.02％.在一定范围内,聚乙二醇、蔗糖、枸橼酸钠、人白蛋白浓度变化对优化方法测定α1-AT生物活性没有影响.结论 以牛胰蛋白酶作为参考标准品的优化α1-AT生物活性测定方法可用于α1-AT制备工艺中的常规质量控制.
目的 驗證優化的α1-抗胰蛋白酶(α1-antitrypsin,α1-AT)生物活性測定方法.方法 用髮色底物法測定α1-AT活性,以牛胰蛋白酶替換凍榦正常人血清來優化先前建立的方法,驗證優化方法的準確性、重複性,併觀察不同物質對優化方法的影響.結果 優化方法在胰蛋白酶質量濃度為0.01～0.05 g/L時線性關繫良好,相關繫數＞0.99.優化方法具有良好的準確性和重複性,噹採用優化方法測定質量濃度為0.01、0.03、0.05 g/L胰蛋白酶時,胰蛋白酶的迴收率分彆為99.33％、94.44％和92.67％,變異繫數分彆為3.79％、1.66％和1.02％.在一定範圍內,聚乙二醇、蔗糖、枸櫞痠鈉、人白蛋白濃度變化對優化方法測定α1-AT生物活性沒有影響.結論 以牛胰蛋白酶作為參攷標準品的優化α1-AT生物活性測定方法可用于α1-AT製備工藝中的常規質量控製.
목적 험증우화적α1-항이단백매(α1-antitrypsin,α1-AT)생물활성측정방법.방법 용발색저물법측정α1-AT활성,이우이단백매체환동간정상인혈청래우화선전건립적방법,험증우화방법적준학성、중복성,병관찰불동물질대우화방법적영향.결과 우화방법재이단백매질량농도위0.01～0.05 g/L시선성관계량호,상관계수＞0.99.우화방법구유량호적준학성화중복성,당채용우화방법측정질량농도위0.01、0.03、0.05 g/L이단백매시,이단백매적회수솔분별위99.33％、94.44％화92.67％,변이계수분별위3.79％、1.66％화1.02％.재일정범위내,취을이순、자당、구연산납、인백단백농도변화대우화방법측정α1-AT생물활성몰유영향.결론 이우이단백매작위삼고표준품적우화α1-AT생물활성측정방법가용우α1-AT제비공예중적상규질량공제.
Objective To validate an optimized method for determination of α1-antitrypsin (o1-AT) activity.Methods The chromogenic substrate method was used for determination of α1-AT activity.The previous established method was optimized by substitution of bovine trypsin for lyophilized normal human serum as reference standard.The accuracy and repeatability of the optimized method were validated,and effects of different substances on the optimized method were observed.Results When trypsin concentrations were in a range of 0.01-0.05 g/L,the optimized method showed a good linearity,and the correlation coefficient was more than 0.99.The optimized method had a good accuracy and repeatability.When 0.01,0.03 and 0.05 g/L trypsin were detected by the optimized method,recovery rates of trypsin were 99.33％,94.44％ and 92.67％,and the variable coefficients were 3.79％,1.66％ and 1.02％,respectively.Concentration changes of polyethylene glycol,sucrose,sodium citrate and human albumin in a certain range did not influence determination of α1-AT activity by the optimized method.Conclusion The optimized method for determination of α1-AT activity with bovine trypsin as reference standard can be used for the routine quality control in o1-AT preparation process.
