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
1期
1-3
,共3页
王玲%潘殊男%张霖阳%夏菊%王宇星%张萍%肖詹蓉
王玲%潘殊男%張霖暘%夏菊%王宇星%張萍%肖詹蓉
왕령%반수남%장림양%하국%왕우성%장평%초첨용
酶联免疫吸附测定%方法%百日咳黏着素
酶聯免疫吸附測定%方法%百日咳黏著素
매련면역흡부측정%방법%백일해점착소
Enzyme-linked immunosorbent assay%Methods%Pertactin
目的 建立定量检测白喉-破伤风-无细胞百日咳疫苗生产过程中黏着素(pertactin,Prn)的方法.方法 纯化的Prn免疫家兔以制备高效价血清抗Prn抗体.辛酸-硫酸铵沉淀法纯化抗Prn多克隆抗体并进行辣根过氧化物酶标记,建立双抗体夹心ELISA法.结果 建立的方法与丝状血凝素和百日咳毒素无交叉反应,特异性较好.该ELISA法在1.25 ～ 80.00μg/L Prn测量区间有最佳线性,相关系数＞0.99.实验内和实验间检测64.0、32.0、16.0 μg/L Prn,变异系数为2.6％～7.7％,回收率为84.9％～ 95.5％,精密度和准确度均符合常规质控要求,因此该法的定量限度为16.0 μg/L.结论 建立的ELISA法可有效检测百日咳疫苗纯化过程中的Prn含量,为组分百日咳疫苗质量控制奠定了重要基础.
目的 建立定量檢測白喉-破傷風-無細胞百日咳疫苗生產過程中黏著素(pertactin,Prn)的方法.方法 純化的Prn免疫傢兔以製備高效價血清抗Prn抗體.辛痠-硫痠銨沉澱法純化抗Prn多剋隆抗體併進行辣根過氧化物酶標記,建立雙抗體夾心ELISA法.結果 建立的方法與絲狀血凝素和百日咳毒素無交扠反應,特異性較好.該ELISA法在1.25 ～ 80.00μg/L Prn測量區間有最佳線性,相關繫數＞0.99.實驗內和實驗間檢測64.0、32.0、16.0 μg/L Prn,變異繫數為2.6％～7.7％,迴收率為84.9％～ 95.5％,精密度和準確度均符閤常規質控要求,因此該法的定量限度為16.0 μg/L.結論 建立的ELISA法可有效檢測百日咳疫苗純化過程中的Prn含量,為組分百日咳疫苗質量控製奠定瞭重要基礎.
목적 건립정량검측백후-파상풍-무세포백일해역묘생산과정중점착소(pertactin,Prn)적방법.방법 순화적Prn면역가토이제비고효개혈청항Prn항체.신산-류산안침정법순화항Prn다극륭항체병진행랄근과양화물매표기,건립쌍항체협심ELISA법.결과 건립적방법여사상혈응소화백일해독소무교차반응,특이성교호.해ELISA법재1.25 ～ 80.00μg/L Prn측량구간유최가선성,상관계수＞0.99.실험내화실험간검측64.0、32.0、16.0 μg/L Prn,변이계수위2.6％～7.7％,회수솔위84.9％～ 95.5％,정밀도화준학도균부합상규질공요구,인차해법적정량한도위16.0 μg/L.결론 건립적ELISA법가유효검측백일해역묘순화과정중적Prn함량,위조분백일해역묘질량공제전정료중요기출.
Objective To establish an ELISA method for quantitative determination of pertactin (Prn) during production of combined diphtheria,tetanus and acellular pertussis vaccine.Methods Rabbits were immuned with purified Prn to prepare high-titer serum antibodies against Prn.The polyclonal antibodies against Prn were purified by octanoic acid-ammonium sulfate precipitation method and labeled with horseradish peroxidase to establish a double antibody sandwich ELISA method.Results The ELISA method had good specificity for Prn and no cross reaction with filamentous hemagglutinin and pertussis toxin.The best linearity of the ELISA method was formed in the range of 1.25-80.00 μg/L (r ＞ 0.99).The coefficient of variation and recover rates of intra-and inter-assay were 2.6％-7.7％ and 84.9％-95.5％ respectively when 64.0,32.0 and 16.0 μg/L standard Prn were detected,and the precision and accuracy both met requirements of quality control.The quantitative limit was 16.0 μg/L.Conclusion The established ELISA method can be applied to detecting Prn during purification of pertussis vaccine and lay the foundation for quality control of Prn.
