Abstract：
A project aimed at establishing replacement batches for the European Pharmacopoeia (Ph. Eur.) Biological Reference Preparation (BRP) Bordetella (B.) pertussis mouse antiserum was started in 2013 under the aegis of the Biological Standardisation Programme (BSP) of the European Directorate for the Quality of Medicines & HealthCare (EDQM). This BRP is used for the immunogenicity assay in mice to assess the potency of acellular pertussis (aP) vaccines as described in Ph. Eur. general method 2.7.16. Assay of pertussis vaccine (acellular). In a preliminary phase of the project (referred to herein as BSP129 phase 1) a hyper-immune serum pool was produced in mice using a combined aP vaccine as immunogen. This pool was used to generate 3 freeze-dried candidate (c) B. pertussis anti-mouse serum BRP batches (cBRP2, cBRP3 and cBRP4). After the pre-qualification that showed their suitability as candidate batches, an international collaborative study (BSP129 phase 2) was carried out in order to standardise these 3 batches against the current BRP1 in terms of anti-PT, -FHA, -PRN and -FIM2/3 antibody contents. For the sake of continuity with the standardisation of BRP1, the corresponding WHO standard (1RR 97/642) was introduced as a second reference for the calibration of the 3 candidate BRPs. Eleven laboratories took part in phase 2. Ten of them performed the ELISA method they use routinely for aP vaccine batch release and one laboratory performed the Multiplex Immunoassay (MIA) as an alternative test. Four participants titrated the antibodies against all 5 pertussis antigens, 5 participants determined the antibody content against 3 antigens (PT, FHA, PRN), one participant titrated the antibodies against PT and FHA antigens and one laboratory determined the antibody content for the PT antigen only. Details of all ELISA methods used were analysed to evaluate their impact on the calibration of the cBRPs. The variability of the results in relation to the nature and methodology of the tests appeared rather limited. Discrepant titres of cBRPs were measured depending on the reference used: the use of the 1RR induced an overestimation (in 8 out of 11 laboratories) and a large inter-laboratory variation in the calculated titres. Regardless of the reference used, equivalency between the calculated titres of cBRP2 and cBRP3 was observed, whilst cBRP4 had systematically lower titres for all antibodies against the 5 acellular pertussis vaccine components. Based on these observations, it was decided to establish the candidate BRP batches against BRP1 and to assign the following potencies based on the mean values determined through centrally calculated results of the calibration assays performed by ELISA in BSP129 phase 2: For cBRP2 and cBRP3 Anti-pertussis toxin: 37 ELISA Units (ELU) per vial Anti-filamentous haemagglutinin: 114 ELU per vial Anti-pertactin: 44 ELU per vial Anti-fimbrial agglutinogens (FIM2/3): 25 ELU per vial For cBRP4 Anti-pertussis toxin: 32 ELU per vial Anti-filamentous haemagglutinin: 98 ELU per vial Anti-pertactin 38 ELU per vial Anti-fimbrial agglutinogens (FIM2/3):23 ELU per vial In February 2018, BRP2, BRP3 and BRP4 were adopted by correspondence by the Ph. Eur. Commission.
摘要：
旨在为欧洲药典建立替代批次的项目（Ph。欧尔.)生物参考制剂(BRP)博德特氏菌(B.）百日咳小鼠抗血清于2013年在欧洲药品质量与医疗保健局（EDQM）的生物标准化计划（BSP）的主持下开始使用。该BRP用于小鼠的免疫原性测定，以评估无细胞百日咳（aP）疫苗的效力。欧尔.一般方法2.7.16。百日咳疫苗(无细胞)的测定。在该项目的初步阶段(本文称为BSP129第1阶段),使用组合aP疫苗作为免疫原,在小鼠中产生超免疫血清池。该库用于产生3个冻干的候选物(c)百日咳博德特氏菌抗小鼠血清BRP批次(cBRP2、cBRP3和cBRP4)。在资格预审表明它们适合作为候选批次后，进行了一项国际合作研究（BSP129第2阶段），以使这3个批次在抗PT方面相对于当前的BRP1标准化，-FHA,-PRN和-FIM2/3抗体含量。为了保持BRP1标准化的连续性，引入了相应的WHO标准（1RR97/642）作为校准3个候选BRP的第二个参考。11个实验室参加了第二阶段。其中10人进行了常规用于aP疫苗批量释放的ELISA方法，一个实验室进行了多重免疫测定（MIA）作为替代测试。四名参与者滴定了针对所有5种百日咳抗原的抗体，5名参与者确定了针对3种抗原的抗体含量（PT，FHA,PRN),一名参与者滴定了针对PT和FHA抗原的抗体,一家实验室仅测定了PT抗原的抗体含量.分析所有使用的ELISA方法的细节以评估它们对cBRP校准的影响。与测试的性质和方法有关的结果的可变性似乎相当有限。根据所使用的参考来测量cBRP的差异滴度：使用1RR会导致高估（11个实验室中的8个），并且计算滴度的实验室间差异很大。无论使用何种参考,观察到cBRP2和cBRP3的计算滴度之间的等效性，而cBRP4针对5种无细胞百日咳疫苗成分的所有抗体的滴度均系统性降低.基于这些观察,决定建立针对BRP1的候选BRP批次，并根据通过在BSP129阶段2中通过ELISA进行的校准测定的集中计算结果确定的平均值来分配以下效力：对于cBRP2和cBRP3抗百日咳毒素：每瓶37ELISA单位（ELU）2月抗丝状血凝素：114ELU每瓶抗百日病凝集素：ELmBRmbrixin/ELmM2采用的EL3欧尔.委员会。
